Compounds &amp; Methods for the Enhanced Degradation of Targeted Proteins &amp; Other Polypeptides by an E3 Ubiquitin Ligase

ABSTRACT

The present invention relates to bifunctional compounds, which find utility as modulators of targeted ubiquitination, especially inhibitors of a variety of polypeptides and other proteins which are degraded and/or otherwise inhibited by bifunctional compounds according to the present invention. In particular, the present invention is directed to compounds, which contain on one end a VHL ligand which binds to the ubiquitin ligase and on the other end a moiety which binds a target protein such that the target protein is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of that protein. The present invention exhibits a broad range of pharmacological activities associated with compounds according to the present invention, consistent with the degradation/inhibition of targeted polypeptides.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation of, and claims priority to,U.S. patent application Ser. No. 14/371,956, filed Jul. 11, 2014, whichis a 35 U.S.C. § 371 national phase application of, and claims priorityto, PCT Application No. PCT/US2013/021136, filed Jan. 11, 2013, whichclaims priority under 35 U.S.C. § 119(e) to U.S. Provisional PatentApplication No. 61/585,769, filed Jan. 12, 2012, all of whichapplications are incorporated herein by reference in their entireties.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under AI084140 awardedby National Institutes of Health. The government has certain rights inthe invention.

FIELD OF THE INVENTION

The present invention relates to bifunctional compounds, which findutility as modulators of targeted ubiquitination, especially inhibitorsof a variety of polypeptides and other proteins, which are degradedand/or otherwise inhibited by bifunctional compounds according to thepresent invention. In particular, the present invention is directed tocompounds, which contain on one end a VHL ligand, which binds to the VHLE3 ubiquitin ligase (defined as a ubiquitin ligand binding moiety or

group) and on the other end a moiety, which binds a target protein(defined as a protein/polypeptide targeting moiety or

group) such that the target protein/polypeptide is placed in proximityto the ubiquitin ligase to effect degradation (and inhibition) of thatprotein. The present invention exhibits a broad range of pharmacologicalactivities associated with compounds according to the present invention,consistent with the degradation/inhibition of targeted polypeptides.

BACKGROUND OF THE INVENTION

E3 ubiquitin ligases (of which over 600 are known in humans)¹ confersubstrate specificity for ubiquitination and are more attractivetherapeutic targets than general proteasome inhibitors^(3,4) due totheir specificity for certain protein substrates. Although thedevelopment of ligands of E3 ligase has proven challenging, in part dueto the fact that they must disrupt protein-protein interactions⁵ recentdevelopments have provided specific ligands which bind to these ligases.Protein-protein interaction interactions are notoriously difficult totarget using small molecules due to their large contact surfaces and theshallow grooves or flat interfaces involved. Conversely, most smallmolecule drugs bind enzymes or receptors in tight and well-definedpockets.⁶ Since the discovery of nutlins, the first small molecule E3ligase inhibitors,⁷ additional compounds have been reported that targetInhibitors of Apoptosis Proteins (IAPs),^(8,9) SCF^(Met30),¹⁰ andSCF^(Cdc4);¹¹ however, the field remains underdeveloped.

One E3 ligase with exciting therapeutic potential is the vonHippel-Lindau (VHL) tumor suppressor, the substrate recognition subunitof the E3 ligase complex VCB, which also consists of elongins B and C,Cul2 and Rbx1.¹² The primary substrate of VHL is Hypoxia InducibleFactor 1α (HIF-1α), a transcription factor that upregulates genes suchas the pro-angiogenic growth factor VEGF and the red blood cell inducingcytokine erythropoietin in response to low oxygen levels. While HIF-1αis constitutively expressed, its intracellular levels are kept very lowunder normoxic conditions via its hydroxylation by prolyl hydroxylasedomain (PHD) proteins and subsequent VHL-mediated ubiquitination (FIG.1).

Using rational design, we have generated the first small moleculeligands of Von Hippel Lindau (VHL), the substrate recognition subunit ofthe E3 ligase VCB, an important target in cancer, chronic anemia andischemia.² We have also obtained crystal structures of VHL with our mostpotent ligand, 15, confirming that the compound mimics the binding modeof the transcription factor HIF-1α, the major substrate of VHL.

From earlier biochemical and structural studies of a hydroxylated HIFpeptide bound to VHL, it became clear that hydroxyproline played animportant role in mediating this protein:protein interaction. As aconsequence of that work, the present inventors developed a hydroxylatedHIF peptide:VHL fluorescence polarization (FP) binding assay with whichthey assayed >120 compounds possessing the central hydroxyprolineresidue flanked by non-peptidic moieties. Further to that research, theinventors now have developed co-crystal structures of VHL complexed withseven of the top compounds. Analysis of these ligand bound structures isdriving the design/synthesis of the next generation of VHL ligands,which are linked with protein binding moieties to produce bifunctionalcompounds according to the present invention.

A principal rationale for the present invention is the need for a smallmolecule E3 ligase ligand for our PROTAC (Proteolysis Targeting Chimera)technology. This technology brings targeted proteins/polypeptides to E3ligases for ubiquitination and subsequent proteasomal degradation. Inseveral proof-of-concept experiments, the present inventors demonstratedthe utility of this approach using the short peptide sequence from HIFthat binds VHL. In order to make a more ‘drug-like’ PROTAC, theinventors have replaced the HIF peptide with a ‘small molecule’ VHLligand, thus providing a means to recruit proteins to E3 ligases forubiquitination and degradation, to the endpoint of providing therapiesbased upon this protein degradation.

OBJECTS OF THE INVENTION

It is an object of the invention to provide compounds which function torecruit endogenous proteins to E3 Ubiquitin Ligase for degradation.

It is an additional object of the invention to provide compounds whichmodulate protein degradation in a patient or subject and can be used fortreating disease states or conditions which are modulated through thedegraded protein.

It is another object of the invention to provide pharmaceuticalcompositions based upon the above-described modulators, especiallyincluding inhibitors for therapeutic treatment of a patient or subject,preferably including a human patient or subject.

It is also an object of the invention to provide methods for determiningprotein binding moieties which bind to proteins of interest.

It is yet another object of the invention to provide methods foridentifying endogenous proteins in a biological system, especiallyincluding a human system, which bind to protein binding moieties incompounds according to the present invention.

It is still another object of the invention to provide methods foridentifying the effects of the degradation of proteins of interest in abiological system using compounds according to the present invention.

It is still another aspect of the invention to provide methods fortreating patients where the degradation of a targeted protein willproduce an intended therapeutic effect.

It is another object of the invention to provide compounds andcompositions which may be used in a first medical application.

It is yet another aspect of the invention to provide compounds and/orcompositons which are used for treating patients where the degradationof a targeted protein will produce an intended therapeutic effect.

Any one or more of these and/or other objects of the invention may bereadily gleaned from a routine scrutiny of the description of theinvention which follows.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows (A) HIF-1α accumulation leads to the transcriptionalupregulation of genes involved in the hypoxic response, such aserythropoietin and VEGF. (B) Under normoxic conditions, HIF-1α ishydroxylated, recognized by VHL, ubiquitinated and degraded by theproteasome, preventing transcriptional upregulation.

FIG. 2. WaterLOGSY NMR spectroscopy shows binding of 3, but not L-Hyp orNAc-Hyp-NMe to VHL.

FIG. 3 shows a pictorial representation shows the key interactionsbetween 15 and VHL.

FIG. 4 shows the 2.9 Å co-crystal structure of 15 (lightest graycarbons) bound to VHL indicates that its binding mimics that of theHIF-1α peptide (light gray carbons, pdb 1LM8¹⁷)

FIG. 5 shows the crystal structures of V₅₄BC apo (A) and in complex with15 (B). Electron density (2F_(o)−F_(c)) superimposed around Hyp bindingsite residues (sticks, yellow carbons) and conserved water molecules(red dots), and 15 (sticks, cyan carbons) are shown in blue and arecontoured at 1.20. The protein surface is shown in green at 50%transparency.

FIGS. 6-12 (each A and B) show the activity of individual compoundsaccording to the present invention in the described VHLpolarization/displacement assay. Compounds according to the presentinvention are indicated with numerals at the top of each graph. Controlcompound is presented in FIG. 15 B and served as minimum polarization(maximum displacement) for comparison purposes. The percent inhibitionas presented was determined by normalizing to maximum and minimumpolarization, and graphed against the log [VL]. IC₅₀ values weredetermined using Prism 5 for each replicate (n=9), where were thenaveraged to determine the average IC₅₀ and the standard of error of themean (SEM).

FIG. 13 (along with Table 2—Affinity Table) shows numerous exemplarycompounds according to the present invention.

FIG. 14 shows numerous preferred compounds from Table 2 according to thepresent invention.

FIG. 15 shows a further number of compounds according to the presentinvention and their activity. Most compounds are active belowconcentrations of 100 μM.

FIG. 16 shows numerous preferred compounds from FIG. 15 according to thepresent invention.

FIG. 17 shows eight particularly preferred compounds from FIG. 15according to the present invention.

FIG. 18 shows six preferred compounds according to the present inventionwhich contain estrogen binding protein targeting moieties linked topreferred ubiquitin ligand binding moieties.

FIG. 19 shows a genus of preferred compounds according to the presentinvention.

BRIEF DESCRIPTION OF THE INVENTION

The present invention is based on the discovery that an ubiquitinpathway protein ubiquitinates any target protein once the ubiquitinpathway protein and the target protein are placed in proximity by achimeric construct that binds the ubiquitin pathway protein and thetarget protein. Accordingly the present invention provides a compositionthat results in the ubiquitination of a chosen target protein. Thepresent invention also provides a library of compositions and the usethereof.

In one embodiment, the present invention provides a composition usefulfor regulating protein activity. The composition comprises a ubiquitinpathway protein binding moiety (preferably for an E3 ubiquitin ligase,alone or in complex with an E2 ubiquitin conjugating enzyme which isresponsible for the transfer of ubiquitin to targeted proteins)according to a defined chemical structure and a protein targeting moietywhich are linked together, preferably through a linker, wherein theubiquitin pathway protein binding moiety recognizes an ubiquitin pathwayprotein and the targeting moiety recognizes a target protein and whereinthe ubiquitin pathway protein binding moiety is coupled to the targetingmoiety.

In another embodiment, the present invention provides a library ofcompounds. The library comprises more than one compound wherein eachcomposition has a formula of A-B, wherein A is a ubiquitin pathwayprotein binding moiety (preferably, an E3 ubiquitin ligase moiety asotherwise disclosed herein) and B is a protein binding member of amolecular library, wherein A is coupled (preferably, through a linkermoiety) to B, and wherein the ubiquitin pathway protein binding moietyrecognizes an ubiquitin pathway protein, in particular, an E3 ubiquitinligase. In a particular embodiment, the library contains a specificubiquitination recognition peptide of VHL for an E3 ubiquitin ligase(ubiquitin pathway protein binding moiety as otherwise disclosed herein)with random target protein binding elements (e.g., a chemical compoundlibrary). As such, the target protein is not determined in advance andthe method can be used to determine the activity of a putative proteinbinding element and its pharmacological value as a target upondegradation by ubiquitin ligase.

In still another embodiment, the present invention provides a method ofscreening a library of the present invention to identify a compoundcontaining a targeting moiety, which recognizes a target proteinassociated with a predetermined function of a cell. The method comprisesincubating a cell with a pool of entities from the library; monitoringthe predetermined function of the cell; identifying a pool of entitiesthat change the predetermined function of the cell; incubating the cellwith a composition from the identified pool of entities; monitoring thepredetermined function of the cell; and identifying a composition thatchanges the predetermined function of the cell, wherein the identifiedcomposition contains a targeting moiety which recognizes a targetprotein associated with the predetermined function.

In another embodiment, the present invention provides a method ofscreening a library of the present invention to identify a compositioncontaining a targeting moiety, which recognizes a target proteinassociated with a predetermined function of a cell. The method comprisesincubating a cell with each composition from the library; monitoring thepredetermined function of the cell; identifying a composition thatchanges the predetermined function of the cell; wherein the identifiedcomposition contains a targeting moiety, which recognizes a targetprotein associated with the predetermined function.

In still another embodiment, the present invention provides a method ofidentifying a target protein associated with a predetermined function ofa cell. The method comprises incubating a cell with a composition fromthe library of the present invention; monitoring the predeterminedfunction of the cell; identifying a composition that changes thepredetermined function of the cell; identifying a target protein thatbinds to the identified composition, wherein the target protein isassociated with the predetermined function of the cell.

In yet another embodiment, the present invention provides a method ofidentifying a target protein associated with a predetermined function ofa cell. The method comprises incubating a cell with a pool of entitiesfrom the library of the present invention; monitoring the predeterminedfunction of the cell; identifying a pool of entities that change thepredetermined function of the cell; incubating the cell with acomposition from the identified pool of entities; monitoring thepredetermined function of the cell; identifying a composition thatchanges the predetermined function of the cell; and identifying a targetprotein that binds to the identified composition, wherein the targetprotein is associated with the predetermined function of the cell.

In yet another embodiment, the present invention provides a method ofubiquitinating/degrading a target protein in a cell. The methodcomprises administering a bifunctional composition comprising anubiquitin pathway protein binding moiety and a targeting moiety,preferably linked through a linker moiety, as otherwise describedherein, wherein the ubiquitin pathway protein binding moiety is coupledto the targeting moiety and wherein the ubiquitin pathway proteinbinding moiety recognizes a ubiquitin pathway protein (e.g., anubiquitin ligase, preferably an E3 ubiquitin ligase) and the targetingmoiety recognizes the target protein such that degradation of the targetprotein will occur when the target protein is placed in proximity to theubiquitin ligase, thus resulting in degradation/inhibition of theeffects of the target protein and the control of protein levels. Thecontrol of protein levels afforded by the present invention providestreatment of a disease state or condition, which is modulated throughthe target protein by lowering the level of that protein in the cells ofa patient.

In another embodiment, the present invention is directed to a method oftreating a patient in need for a disease state or condition modulatedthrough a protein where the degradation of that protein will produce atherapeutic effect in that patient, the method comprising administeringto a patient in need an effective amount of a compound according to thepresent invention, optionally in combination with another bioactiveagent. The disease state or condition may be a disease caused by amicrobial agent or other exogenous agent such as a virus, bacteria,fungus, protozoa or other microbe or may be a disease state, which iscaused by overexpression of a protein, which leads to a disease stateand/or condition.

In one embodiment, the present invention is directed to a compoundaccording to the structure:

Where L is a linker group and

is a ubiquitin ligase binding moiety, wherein said linker group isoptionally further linked to a

group.

In another embodiment, the present invention is directed to a compoundwhich comprises a

group according to the general structure:

Where

is an ubiquitin ligase binding moiety, preferably a ligand, which bindsan ubiquitin ligase, preferably an E3 ubiquitin ligase;

is a chemical moiety (protein targeting moiety), which binds to a targetprotein or polypeptide, which is ubiquitinated by an ubiquitin ligaseand is chemically linked directly to the

group or through a linker moiety L, or

is alternatively a

group which also an ubiquitin ligase binding moiety, which may be thesame or different than the

group and is linked directly to the

group directly or through the linker moiety; andL is a linker moiety which may be present or absent and which chemically(covalently) links

to

,Or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvateor polymorph thereof.

In certain aspects of the invention, where

is a

group, the compound resembles a dimeric compound where both ends of thecompound comprise an ubiquitin ligase binding moiety as otherwisedescribed herein.

In preferred aspects of the invention,

and where present,

, are each independently a group according to the chemical structure:

Where R^(1′) is an optionally substituted C₁-C₆ alkyl group, anoptionally substituted —(CH₂)_(n)OH, an optionally substituted—(CH₂)_(n)SH, an optionally substituted (CH₂)_(n)—O—(C₁-C₆)alkyl group,an optionally substituted (CH₂)_(n)—WCOCW—(C₀-C₆)alkyl group containingan epoxide moiety WCOCW where each W is independently H or a C₁-C₃ alkylgroup, an optionally substituted —(CH₂)_(n)COOH, an optionallysubstituted —(CH₂)_(n)C(O)—(C₁-C₆ alkyl), an optionally substituted—(CH₂)_(n)NHC(O)—R₁, an optionally substituted —(CH₂)_(n)C(O)—NR₁R₂, anoptionally substituted —(CH₂)_(n)OC(O)—NR₁R₂, —(CH₂O)_(n)H, anoptionally substituted —(CH₂)_(n)OC(O)—(C₁-C₆ alkyl), an optionallysubstituted —(CH₂)_(n)C(O)—O—(C₁-C₆ alkyl), an optionally substituted—(CH₂O)_(n)COOH, an optionally substituted —(OCH₂)_(n)O—(C₁-C₆ alkyl),an optionally substituted —(CH₂O)_(n)C(O)—(C₁-C₆ alkyl), an optionallysubstituted —(OCH₂)_(n)NHC(O)—R₁, an optionally substituted—(CH₂O)_(n)C(O)—NR₁R₂, —(CH₂CH₂O)_(n)H, an optionally substituted—(CH₂CH₂O)_(n)COOH, an optionally substituted —(OCH₂CH₂)_(n)O—(C₁-C₆alkyl), an optionally substituted —(CH₂CH₂O)_(n)C(O)—(C₁-C₆ alkyl), anoptionally substituted —(OCH₂CH₂)_(n)NHC(O)—R₁, an optionallysubstituted —(CH₂CH₂O)_(n)C(O)—NR₁R₂, an optionally substituted—SO₂R_(S), an optionally substituted S(O)R_(S), NO₂, CN or halogen (F,Cl, Br, I, preferably F or Cl);R₁ and R₂ are each independently H or a C₁-C₆ alkyl group which may beoptionally substituted with one or two hydroxyl groups or up to threehalogen groups (preferably fluorine);R_(S) is a C₁-C₆ alkyl group, an optionally substituted aryl, heteroarylor heterocycle group or a —(CH₂)_(m)NR₁R₂ group,X and X′ are each independently C═O, C═S, —S(O), S(O)₂, (preferably Xand X′ are both C═O);R^(2′) is an optionally substituted—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)alkyl group, an optionallysubstituted —(CH₂)_(n)—(C═O)_(u)(NR)_(v)(SO₂)_(w)NR_(1N)R_(2N) group, anoptionally substituted —(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl, anoptionally substituted—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Heteroaryl, an optionallysubstituted —(CH₂)_(n)—(C═O)_(v)NR₁(SO₂)_(w)-Heterocycle, an optionallysubstituted —NR¹—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-alkyl, anoptionally substituted—NR¹—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), an optionallysubstituted —NR¹—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR₁C(O)R_(1N), anoptionally substituted —NR¹—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl,an optionally substituted—NR¹—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Heteroaryl or an optionallysubstituted —NR¹—(CH₂)_(n)—(C═O)_(v)NR₁(SO₂)_(w)-Heterocycle, anoptionally substituted —X^(R2′)-alkyl group; an optionally substituted—X^(R2′)— Aryl group; an optionally substituted —X^(R2′)— Heteroarylgroup; an optionally substituted —X^(R2′)— Heterocycle group; anoptionally substituted; R^(3′) is an optionally substituted alkyl, anoptionally substituted —(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-alkyl, anoptionally substituted—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), an optionallysubstituted —(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR₁C(O)R_(1N), anoptionally substituted —(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—C(O)NR₁R₂,an optionally substituted —(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl, anoptionally substituted —(CH₂)_(n)—C(O)(NR₁)(SO₂)_(w)-Heteroaryl, anoptionally substituted —(CH₂)_(n)—C(O)(NR₁)(SO₂)_(w)-Heterocycle, anoptionally substituted —NR¹—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-alkyl,an optionally substituted—NR¹—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), an optionallysubstituted —NR¹—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR₁C(O)R_(1N), anoptionally substituted —NR¹—(CH₂)_(n)—C(O)(NR₁)_(v)(SO₂)_(w)-Aryl, anoptionally substituted —NR¹—(CH₂)_(n)—C(O)(NR₁)_(v)(SO₂)_(w)-Heteroaryl,an optionally substituted—NR¹—(CH₂)_(n)—C(O)(NR₁)_(v)(SO₂)_(w)-Heterocycle, an optionallysubstituted —O—(CH₂)_(n)—(C═O)(NR₁)_(v)(SO₂)_(w)-alkyl, an optionallysubstituted —O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), anoptionally substituted—O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)—NR₁C(O)R_(1N), an optionallysubstituted —O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl, an optionallysubstituted —O—(CH₂)_(n)—(C═O)(NR₁)_(v)(SO₂)_(w)-Heteroaryl or anoptionally substituted—O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Heterocycle;—(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)-alkyl group, an optionallysubstituted —(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)-Aryl group, anoptionally substituted —(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)-Heteroarylgroup, an optionally substituted—(CH₂)_(n)—(V)_(n)—(CH₂)_(n)—(V)_(n′)-Heterocycle group, an optionallysubstituted —(CH₂)_(n)—N(R₁)(C═O)_(m′)—(V)_(n′)-alkyl group, anoptionally substituted —(CH₂)_(n)—N(R₁)(C═O)_(m′)—(V)_(n′)-Aryl group,an optionally substituted —(CH₂)_(n)—N(R₁)(C═O)_(m′)—(V)_(n′)-Heteroarylgroup, an optionally substituted—(CH₂)_(n)—N(R₁)(C═O)_(m′)—(V)_(n′)-Heterocycle group, an optionallysubstituted —X^(R3′)-alkyl group; an optionally substituted—X^(R3′)-Aryl group; an optionally substituted —X^(R3′)-Heteroarylgroup; an optionally substituted —X^(R3′)-Heterocycle group; anoptionally substituted;Where R_(1N) and R_(2N) are each independently H, C₁-C₆ alkyl which isoptionally substituted with one or two hydroxyl groups and up to threehalogen groups or an optionally substituted —(CH₂)_(n)-Aryl,—(CH₂)_(n)-Heteroaryl or —(CH₂)_(n)-Heterocycle group;

V is O, S or NR₁;

R₁ is the same as above;R¹ and R_(1′) are each independently H or a C₁-C₃ alkyl group;X^(R2′) and X^(R3′) are each independently an optionally substituted—CH₂)_(n)—, —CH₂)_(n)—CH(X_(v))═CH(X_(v))— (cis or trans),—CH₂)_(n)—CH≡CH—, —(CH₂CH₂O)_(n)— or a C₃-C₆ cycloalkyl group, whereX_(v) is H, a halo or a C₁-C₃ alkyl group which is optionallysubstituted;Each m is independently 0, 1, 2, 3, 4, 5, 6;Each m′ is independently 0 or 1;Each n is independently 0, 1, 2, 3, 4, 5, 6;Each n′ is independently 0 or 1;Each u is independently 0 or 1;Each v is independently 0 or 1;Each w is independently 0 or 1; andwherein any one or more of R^(1′), R^(2′), R^(3′), X and X′ of

is modified to be covalently bonded to the

group through a linker group when

is not,

, or when

is

, any one or more of R^(1′), R^(2′), R^(3′), X and X′ of each of

and

are modified to be covalently bonded to each other directly or through alinker group, ora pharmaceutically acceptable salt, stereoisomer, solvate orpolymorphthereof.

In alternative aspects of the present invention

and when present,

, are each independently a group according to the chemical structure:

Wherein each of R^(1′), R^(2′) and R^(3′) are the same as above and X isC═O, C═S, —S(O) group or a S(O)₂ group, more preferably a C═O group, andwherein any one or more of R^(1′), R^(2′) and R^(3′) are modified tobind a linker group to which is further covalently bonded to the

group when

is not

, or when

is

, any one or more of R^(1′), R^(2′), R^(3′) of each of

and

are modified to be covalently bonded to each other directly or through alinker group, or a pharmaceutically acceptable salt, enantiomer,diastereomer, solvate or polymorph thereof.

In still further preferred aspects of the invention,

, and when present,

, are each independently according to the chemical structure:

wherein any one or more of R^(1′), R^(2′) and R^(3′) are modified tobind a linker group to which is further covalently bonded to the

group when

is not

, or when

is

, any one or more of R^(1′), R^(2′), R^(3′) of each of

and

are modified to be covalently bonded to each other directly or through alinker group, or a pharmaceutically acceptable salt, enantiomer,diastereomer, solvate or polymorph thereof.

In further preferred aspects of the invention, R^(1′) is preferably ahydroxyl group or a group which may be metabolized to a hydroxyl orcarboxylic group, such that the compound represents a prodrug form of anactive compound. Exemplary preferred R^(1′) groups include, for example,—(CH₂)_(n)OH, (CH₂)_(n)—O—(C₁-C₆)alkyl group, —(CH₂)_(n)COOH,—(CH₂O)_(n)H, an optionally substituted —(CH₂)_(n)OC(O)—(C₁-C₆ alkyl),or an optionally substituted —(CH₂)_(n)C(O)—O—(C₁-C₆ alkyl), wherein nis 0 or 1. Where R^(1′) is or contains a carboxylic acid group, ahydroxyl group or an amine group, the hydroxyl group, carboxylic acidgroup or amine (each of which may be optionally substituted), may befurther chemically modified to provide a covalent link to a linker groupto which the

group (including a

group) is bonded.

X and X′, where present, are preferably a C═O, C═S, —S(O) group or aS(O)₂ group, more preferably a C═O group.

R^(2′) is preferably an optionally substituted —NR¹-T-Aryl, anoptionally substituted —NR¹-T-Heteroaryl group or an optionallysubstituted —NR¹-T-Heterocycle, where R¹ is H or CH₃, preferably H and Tis an optionally substituted —(CH₂)_(n)— group, wherein each one of themethylene groups may be optionally substituted with one or twosubstituents, preferably selected from halogen, an amino acid sidechainas otherwise described herein or a C₁-C₃ alkyl group, preferably one ortwo methyl groups, which may be optionally substituted; and n is 0 to 6,often 0, 1, 2 or 3, preferably 0 or 1. Alternatively, T may also be a—(CH₂O)_(n)— group, a —(OCH₂)_(n)— group, a —(CH₂CH₂O)_(n)— group, a—(OCH₂CH₂)_(n)— group, all of which groups are optionally substituted.

Preferred Aryl groups for R^(2′) include optionally substituted phenylor naphthyl groups, preferably phenyl groups, wherein the phenyl groupis optionally substituted with a linker group to which is attached a

group (including a

group), a halogen (preferably F or Cl), an amine, monoalkyl- or dialkylamine (preferably, dimethylamine), F, Cl, OH, COOH, C₁-C₆ alkyl,preferably CH₃, CF₃, OMe, OCF₃, NO₂, or CN group (each of which may besubstituted in ortho-, meta- and/or para-positions of the phenyl ring,preferably para-), an optionally substituted phenyl group (the phenylgroup itself is preferably substituted with a linker group attached to a

group, including a

group), and/or at least one of F, Cl, OH, COOH, CH₃, CF₃, OMe, OCF₃,NO₂, or CN group (in ortho-, meta- and/or para-positions of the phenylring, preferably para-), a naphthyl group, which may be optionallysubstituted, an optionally substituted heteroaryl, preferably anoptionally substituted isoxazole including a methylsubstitutedisoxazole, an optionally substituted oxazole including amethylsubstituted oxazole, an optionally substituted thiazole includinga methyl substituted thiazole, an optionally substituted isothiazoleincluding a methyl substituted isothiazole, an optionally substitutedpyrrole including a methylsubstituted pyrrole, an optionally substitutedimidazole including a methylimidazole, an optionally substitutedbenzimidazole or methoxybenzylimidazole, an optionally substitutedoximidazole or methyloximidazole, an optionally substituted diazolegroup, including a methyldiazole group, an optionally substitutedtriazole group, including a methylsubstituted triazole group, anoptionally substituted pyridine group, including a halo- (preferably, F)or methylsubstitutedpyridine group or an oxapyridine group (where thepyridine group is linked to the phenyl group by an oxygen), anoptionally substituted furan, an optionally substituted benzofuran, anoptionally substituted dihydrobenzofuran, an optionally substitutedindole, indolizine or azaindolizine (2, 3, or 4-azaindolizine), anoptionally substituted quinoline, an optionally substituted groupaccording to the chemical structure:

Where S^(c) is CHR^(SS), NR^(URE), or O;R^(HET) is H, CN, NO₂, halo (preferably Cl or F), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups (e.g. CF₃), optionally substituted O(C₁-C₆alkyl) (preferably substituted with one or two hydroxyl groups or up tothree halo groups) or an optionally substituted acetylenic group—C≡C—R_(a) where R_(a) is H or a C₁-C₆ alkyl group (preferably C₁-C₃alkyl);R^(SS) is H, CN, NO₂, halo (preferably F or Cl), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups), optionally substituted O—(C₁-C₆ alkyl)(preferably substituted with one or two hydroxyl groups or up to threehalo groups) or an optionally substituted —C(O)(C₁-C₆ alkyl) (preferablysubstituted with one or two hydroxyl groups or up to three halo groups);R^(URE) is H, a C₁-C₆ alkyl (preferably H or C₁-C₃ alkyl) or a—C(O)(C₁-C₆ alkyl) each of which groups is optionally substituted withone or two hydroxyl groups or up to three halogen, preferably fluorinegroups, or an optionally substituted phenyl group, an optionallysubstituted heteroaryl, or an optionally substituted heterocycle,preferably for example piperidine, morpholine, pyrrolidine,tetrahydrofuran);R^(PRO) is H, optionally substituted C₁-C₆ alkyl or an optionallysubstituted aryl (phenyl or napthyl), heteroaryl or heterocyclic groupselected from the group consisting of oxazole, isoxazole, thiazole,isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine,furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,quinoline, (each preferably substituted with a C₁-C₃ alkyl group,preferably methyl or a halo group, preferably F or Cl), benzofuran,indole, indolizine, azaindolizine;R^(PRO1) and R^(PRO2) are each independently H, an optionallysubstituted C₁-C₃ alkyl group or together form a keto group; andeach n is independently 0, 1, 2, 3, 4, 5, or 6 (preferably 0 or 1), oran optionally substituted heterocycle, preferably tetrahydrofuran,tetrahydrothiene, piperidine, piperazine or morpholine (each of whichgroups when substituted, are preferably substituted with a methyl orhalo (F, Br, Cl),each of which groups may be optionally substituted with a linker groupto which is attached a

group (including a

group).

In certain preferred aspects,

is a

group,Where R^(PRO) and n are the same as above.

Preferred heteroaryl groups for R^(2′) include an optionally substitutedquinoline (which may be attached to the pharmacophore or substituted onany carbon atom within the quinoline ring), an optionally substitutedindole, an optionally substituted indolizine, an optionally substitutedazaindolizine, an optionally substituted benzofuran, including anoptionally substituted benzofuran, an optionally substituted isoxazole,an optionally substituted thiazole, an optionally substitutedisothiazole, an optionally substituted thiophene, an optionallysubstituted pyridine (2-, 3, or 4-pyridine), an optionally substitutedimidazole, an optionally substituted pyrrole, an optionally substituteddiazole, an optionally substituted triazole, a tetrazole, an optionallysubstituted oximidazole, or a group according to the chemical structure:

Where S^(c) is CHR^(SS), NR^(URE), or O;R^(HET) is H, CN, NO₂, halo (preferably Cl or F), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups (e.g. CF₃), optionally substituted O(C₁-C₆alkyl) (preferably substituted with one or two hydroxyl groups or up tothree halo groups) or an optionally substituted acetylenic group—C≡C—R_(a) where R_(a) is H or a C₁-C₆ alkyl group (preferably C₁-C₃alkyl);R^(SS) is H, CN, NO₂, halo (preferably F or Cl), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups), optionally substituted O—(C₁-C₆ alkyl)(preferably substituted with one or two hydroxyl groups or up to threehalo groups) or an optionally substituted —C(O)(C₁-C₆ alkyl) (preferablysubstituted with one or two hydroxyl groups or up to three halo groups);R^(URE) is H, a C₁-C₆ alkyl (preferably H or C₁-C₃ alkyl) or a—C(O)(C₁-C₆ alkyl), each of which groups is optionally substituted withone or two hydroxyl groups or up to three halogen, preferably fluorinegroups, or an optionally substituted heterocycle, for examplepiperidine, morpholine, pyrrolidine, tetrahydrofuran,tetrahydrothiophene, piperidine, piperazine, each of which is optionallysubstituted, andY^(C) is N or C—R^(YC), where R^(YC) is H, OH, CN, NO₂, halo (preferablyCl or F), optionally substituted C₁-C₆ alkyl (preferably substitutedwith one or two hydroxyl groups or up to three halo groups (e.g. CF₃),optionally substituted O(C₁-C₆ alkyl) (preferably substituted with oneor two hydroxyl groups or up to three halo groups) or an optionallysubstituted acetylenic group —C≡C—R_(a) where R_(a) is H or a C₁-C₆alkyl group (preferably C₁-C₃ alkyl), each of which groups may beoptionally substituted with a linker group to which is attached a

group (including a

group);

Preferred heterocycle groups for R^(2′) include tetrahydrofuran,tetrahydrothiene, tetrahydroquinoline, piperidine, piperazine,pyrrolidine, morpholine, oxane or thiane, each of which groups may beoptionally substituted, or a group according to the chemical structure:

Preferably, a

group,Where R^(PRO) is H, optionally substituted C₁-C₆ alkyl or an optionallysubstituted aryl, heteroaryl or heterocyclic group;R^(PRO1) and R^(PRO2) are each independently H, an optionally subsitutedC₁-C₃ alkyl group or together form a keto group andEach n is independently 0, 1, 2, 3, 4, 5, or 6 (often 0 or 1), each ofwhich groups may be optionally substituted with a linker group to whichis attached a

group (including a

group).

Preferred R^(2′) substituents for use in the present invention alsoinclude specifically (and without limitation to the specific compounddisclosed) the R^(2′) substituents which are found in the identifiedcompounds disclosed herein (which includes the specific compounds whichare disclosed in the present specification, and the figures which areattached hereto). Each of these R^(2′) substituents may be used inconjunction with any number of R^(3′) substituents which are alsodisclosed herein.

R^(3′) is preferably an optionally substituted -T-Aryl, an optionallysubstituted -T-Heteroaryl, an optionally substituted -T-Heterocycle, anoptionally substituted —NR¹-T-Aryl, an optionally substituted—NR¹-T-Heteroaryl or an optionally substituted —NR¹-T-Heterocycle, whereR¹ is H or a C₁-C₃ alkyl group, preferably H or CH₃, T is an optionallysubstituted —(CH₂)_(n)— group, wherein each one of the methylene groupsmay be optionally substituted with one or two substituents, preferablyselected from halogen, a C₁-C₃ alkyl group or the sidechain of an aminoacid as otherwise described herein, preferably methyl, which may beoptionally substituted; and n is 0 to 6, often 0, 1, 2, or 3 preferably0 or 1. Alternatively, T may also be a —(CH₂O)_(n)— group, a—(OCH₂)_(n)— group, a —(CH₂CH₂O)_(n)— group, a —(OCH₂CH₂)_(n)— group,each of which groups is optionally substituted.

Preferred aryl groups for R³ include optionally substituted phenyl ornaphthyl groups, preferably phenyl groups, wherein the phenyl ornaphthyl group s optionally substituted with a linker group to which isattached a

group (including a

group) and/or a halogen (preferably F or Cl), an amine, monoalkyl- ordialkyl amine (preferably, dimethylamine), an amido group (preferably a—(CH₂)_(m)—NR₁C(O)R₂ group where m, R₁ and R₂ are the same as above), ahalo (often F or Cl), OH, CH₃, CF₃, OMe, OCF₃, NO₂, CN or a S(O)₂R_(S)group (R_(S) is a a C₁-C₆ alkyl group, an optionally substituted aryl,heteroaryl or heterocycle group or a —(CH₂)_(m)NR₁R₂ group), each ofwhich may be substituted in ortho-, meta- and/or para-positions of thephenyl ring, preferably para-), or an Aryl (preferably phenyl),Heteroaryl or Heterocycle. Preferably said substituent phenyl group isan optionally substituted phenyl group (i.e., the substituent phenylgroup itself is preferably substituted with at least one of F, Cl, OH,SH, COOH, CH₃, CF₃, OMe, OCF₃, NO₂, CN or a linker group to which isattached a

group (including a

group), wherein the substitution occurs in ortho-, meta- and/orpara-positions of the phenyl ring, preferably para-), a naphthyl group,which may be optionally substituted including as described above, anoptionally substituted heteroaryl (preferably an optionally substitutedisoxazole including a methylsubstituted isoxazole, an optionallysubstituted oxazole including a methylsubstituted oxazole, an optionallysubstituted thiazole including a methyl substituted thiazole, anoptionally substituted pyrrole including a methylsubstituted pyrrole, anoptionally substituted imidazole including a methylimidazole, abenzylimidazole or methoxybenzylimidazole, an oximidazole ormethyloximidazole, an optionally substituted diazole group, including amethyldiazole group, an optionally substituted triazole group, includinga methylsubstituted triazole group, a pyridine group, including a halo-(preferably, F) or methylsubstitutedpyridine group or an oxapyridinegroup (where the pyridine group is linked to the phenyl group by anoxygen) or an optionally substituted heterocycle (tetrahydrofuran,tetrahydrothiophene, pyrrolidine, piperidine, morpholine, piperazine,tetrahydroquinoline, oxane or thiane. Each of the aryl, heteroaryl orheterocyclic groups may be optionally substituted with a linker group towhich is attached a

group (including a

group).

Preferred Heteroaryl groups for R³ include an optionally substitutedquinoline (which may be attached to the pharmacophore or substituted onany carbon atom within the quinoline ring), an optionally substitutedindole (including dihydroindole), an optionally substituted indolizine,an optionally substituted azaindolizine (2, 3 or 4-azaindolizine) anoptionally substituted benzimidazole, benzodiazole, benzoxofuran, anoptionally substituted imidazole, an optionally substituted isoxazole,an optionally substituted oxazole (preferably methyl substituted), anoptionally substituted diazole, an optionally substituted triazole, atetrazole, an optionally substituted benzofuran, an optionallysubstituted thiophene, an optionally substituted thiazole (preferablymethyl and/or thiol substituted), an optionally substituted isothiazole,an optionally substituted triazole (preferably a 1,2,3-triazolesubstituted with a methyl group, a triisopropylsilyl group, anoptionally substituted —(CH₂)_(m)—O—C₁-C₆ alkyl group or an optionallysubstituted —(CH₂)_(m)—C(O)—O—C₁-C₆ alkyl group), an optionallysubstituted pyridine (2-, 3, or 4-pyridine) or a group according to thechemical structure:

Where S^(c) is CHR^(SS), NR^(URE), or O;R^(HET) is H, CN, NO₂, halo (preferably Cl or F), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups (e.g. CF₃), optionally substituted O(C₁-C₆alkyl) (preferably substituted with one or two hydroxyl groups or up tothree halo groups) or an optionally substituted acetylenic group—C≡C—R_(a) where R_(a) is H or a C₁-C₆ alkyl group (preferably C₁-C₃alkyl);R^(SS) is H, CN, NO₂, halo (preferably F or Cl), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups), optionally substituted O—(C₁-C₆ alkyl)(preferably substituted with one or two hydroxyl groups or up to threehalo groups) or an optionally substituted —C(O)(C₁-C₆ alkyl) (preferablysubstituted with one or two hydroxyl groups or up to three halo groups);R^(URE) is H, a C₁-C₆ alkyl (preferably H or C₁-C₃ alkyl) or a—C(O)(C₁-C₆ alkyl), each of which groups is optionally substituted withone or two hydroxyl groups or up to three halogen, preferably fluorinegroups, or an optionally substituted heterocycle, for examplepiperidine, morpholine, pyrrolidine, tetrahydrofuran,tetrahydrothiophene, piperidine, piperazine, each of which is optionallysubstituted, andY^(C) is N or C—R^(YC), where R^(YC) is H, OH, CN, NO₂, halo (preferablyCl or F), optionally substituted C₁-C₆ alkyl (preferably substitutedwith one or two hydroxyl groups or up to three halo groups (e.g. CF₃),optionally substituted O(C₁-C₆ alkyl) (preferably substituted with oneor two hydroxyl groups or up to three halo groups) or an optionallysubstituted acetylenic group —C≡C—R_(a) where R_(a) is H or a C₁-C₆alkyl group (preferably C₁-C₃ alkyl). Each of said heteroaryl groups maybe optionally substituted with a linker group to which is attached a

group (including a

group).

Preferred heterocycle groups for R^(3′) include tetrahydroquinoline,piperidine, piperazine, pyrrolidine, morpholine, tetrahydrofuran,tetrahydrothiophene, oxane and thiane, each of which groups may beoptionally substituted or a group according to the chemical structure:

Preferably, a

group, Where R^(PRO) is H, optionally substituted C₁-C₆ alkyl or anoptionally substituted aryl (phenyl or napthyl), heteroaryl orheterocyclic group selected from the group consisting of oxazole,isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole,pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene,dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine,morpholine, quinoline, (each preferably substituted with a C₁-C₃ alkylgroup, preferably methyl or a halo group, preferably F or Cl),benzofuran, indole, indolizine, azaindolizine;R^(PRO1) and R^(PRO2) are each independently H, an optionallysubstituted C₁-C₃ alkyl group or together form a keto group, andEach n is 0, 1, 2, 3, 4, 5, or 6 (preferably 0 or 1), wherein each ofsaid Heteocycle groups may be optionally substituted with a linker groupto which is attached a

group (including a

group).

Preferred R^(3′) substituents for use in the present invention alsoinclude specifically (and without limitation to the specific compounddisclosed) the R^(3′) substituents which are found in the identifiedcompounds disclosed herein (which includes the specific compounds whichare disclosed in the present specification, and the figures which areattached hereto). Each of these R^(3′) substituents may be used inconjunction with any number of R^(2′) substituents which are alsodisclosed herein.

In certain alternative preferred embodiments, R^(2′) is an optionallysubstituted —NR₁—X^(R2′)-alkyl group, —NR₁—X^(R2′)-Aryl group; anoptionally substituted —NR₁—X^(R2′)-HET, an optionally substituted—NR₁—X^(R2′)-Aryl-HET or an optionally substituted—NR₁—X^(R2′)-HET-Aryl,

Where R₁ is H or a C₁-C₃ alkyl group (preferably H);X^(R2′) is an optionally substituted —CH₂)_(n)—,—CH₂)_(n)—CH(X_(v))═CH(X_(v))— (cis or trans), —CH₂)_(n)—CH≡CH—,—(CH₂CH₂O)_(n)— or a C₃-C₆ cycloalkyl group;where X_(v) is H, a halo or a C₁-C₃ alkyl group which is optionallysubstituted with one or two hydroxyl groups or up to three halogengroups;Alkyl is an optionally substituted C1-C₁₀ alkyl (preferably a C₁-C₆alkyl) group (in certain preferred embodiments, the alkyl group isend-capped with a halo group, often a Cl or Br); Aryl is an optionallysubstituted phenyl or naphthyl group (preferably, a phenyl group); andHET is an optionally substituted oxazole, isoxazole, thiazole,isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine,furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,benzofuran, indole, indolizine, azaindolizine, quinoline (whensubstituted, each preferably substituted with a C₁-C₃ alkyl group,preferably methyl or a halo group, preferably F or Cl) or a groupaccording to the chemical structure:

Where S^(c) is CHR^(SS), NR^(URE), or O;R^(HET) is H, CN, NO₂, halo (preferably Cl or F), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups (e.g. CF₃), optionally substituted O(C₁-C₆alkyl) (preferably substituted with one or two hydroxyl groups or up tothree halo groups) or an optionally substituted acetylenic group—C≡C—R_(a) where R_(a) is H or a C₁-C₆ alkyl group (preferably C₁-C₃alkyl);R^(SS) is H, CN, NO₂, halo (preferably F or Cl), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups), optionally substituted O—(C₁-C₆ alkyl)(preferably substituted with one or two hydroxyl groups or up to threehalo groups) or an optionally substituted —C(O)(C₁-C₆ alkyl) (preferablysubstituted with one or two hydroxyl groups or up to three halo groups);R^(URE) is H, a C₁-C₆ alkyl (preferably H or C₁-C₃ alkyl) or a—C(O)(C₁-C₆ alkyl), each of which groups is optionally substituted withone or two hydroxyl groups or up to three halogen, preferably fluorinegroups, or an optionally substituted heterocycle, for examplepiperidine, morpholine, pyrrolidine, tetrahydrofuran,tetrahydrothiophene, piperidine, piperazine, each of which is optionallysubstituted, andY^(C) is N or C—R^(YC), where R^(YC) is H, OH, CN, NO₂, halo (preferablyCl or F), optionally substituted C₁-C₆ alkyl (preferably substitutedwith one or two hydroxyl groups or up to three halo groups (e.g. CF₃),optionally substituted O(C₁-C₆ alkyl) (preferably substituted with oneor two hydroxyl groups or up to three halo groups) or an optionallysubstituted acetylenic group —C≡C—R_(a) where R_(a) is H or a C₁-C₆alkyl group (preferably C₁-C₃ alkyl);R^(PRO) is H, optionally substituted C₁-C₆ alkyl or an optionallysubstituted aryl (phenyl or napthyl), heteroaryl or heterocyclic groupselected from the group consisting of oxazole, isoxazole, thiazole,isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine,furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,quinoline, (each preferably substituted with a C₁-C₃ alkyl group,preferably methyl or a halo group, preferably F or Cl), benzofuran,indole, indolizine, azaindolizine;R^(PRO1) and R^(PRO2) are each independently H, an optionallysubstituted C₁-C₃ alkyl group or together form a keto group, andEach n is independently 0, 1, 2, 3, 4, 5, or 6 (preferably 0 or 1). Eachof said groups may be optionally substituted with a linker group towhich is attached a

group (including a

group).

In certain alternative preferred embodiments of the present invention,R^(3′) is an optionally substituted—(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)—R^(S3′) group, an optionallysubstituted —(CH₂)_(n)—N(R_(1′))(C═O)_(m′)—(V)_(n′)—R^(S3′) group, anoptionally substituted —X^(R3′)-alkyl group, an optionally substituted—X^(R3′)-Aryl group; an optionally substituted —X^(R3′)-HET group, anoptionally substituted —X^(R3′)-Aryl-HET group or an optionallysubstituted —X^(R3′)-HET-Aryl group,

Where R^(S3′) is an optionally substituted alkyl group (C₁-C₁₀,preferably C₁-C₆ alkyl), an optionally substituted Aryl group or a HETgroup;R_(1′) is H or a C₁-C₃ alkyl group (preferably H);

V is O, S or NR_(1′);

X^(R3′) is —(CH₂)_(n)—, —(CH₂CH₂O)_(n)—, —CH₂)_(n)—CH(X_(v))═CH(X_(v))—(cis or trans), —CH₂)_(n)—CH≡CH—, or a C₃-C₆ cycloalkyl group, alloptionally substituted;where X_(v) is H, a halo or a C₁-C₃ alkyl group which is optionallysubstituted with one or two hydroxyl groups or up to three halogengroups;Alkyl is an optionally substituted C₁-C₁₀ alkyl (preferably a C₁-C₆alkyl) group (in certain preferred embodiments, the alkyl group isend-capped with a halo group, often a Cl or Br); Aryl is an optionallysubstituted phenyl or napthyl group (preferably, a phenyl group); andHET is an optionally substituted oxazole, isoxazole, thiazole,isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine,furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,benzofuran, indole, indolizine, azaindolizine, quinoline (whensubstituted, each preferably substituted with a C₁-C₃ alkyl group,preferably methyl or a halo group, preferably F or Cl), or a groupaccording to the chemical structure:

Where S^(c) is CHR^(SS), NR^(URE), or O;R^(HET) is H, CN, NO₂, halo (preferably Cl or F), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups (e.g. CF₃), optionally substituted O(C₁-C₆alkyl) (preferably substituted with one or two hydroxyl groups or up tothree halo groups) or an optionally substituted acetylenic group—C≡C—R_(a) where R_(a) is H or a C₁-C₆ alkyl group (preferably C₁-C₃alkyl);R^(SS) is H, CN, NO₂, halo (preferably F or Cl), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups), optionally substituted O—(C₁-C₆ alkyl)(preferably substituted with one or two hydroxyl groups or up to threehalo groups) or an optionally substituted —C(O)(C₁-C₆ alkyl) (preferablysubstituted with one or two hydroxyl groups or up to three halo groups);R^(URE) is H, a C₁-C₆ alkyl (preferably H or C₁-C₃ alkyl) or a—C(O)(C₀-C₆ alkyl), each of which groups is optionally substituted withone or two hydroxyl groups or up to three halogen, preferably fluorinegroups, or an optionally substituted heterocycle, for examplepiperidine, morpholine, pyrrolidine, tetrahydrofuran,tetrahydrothiophene, piperidine, piperazine, each of which is optionallysubstituted, andY^(C) is N or C—R^(YC), where R^(YC) is H, OH, CN, NO₂, halo (preferablyCl or F), optionally substituted C₁-C₆ alkyl (preferably substitutedwith one or two hydroxyl groups or up to three halo groups (e.g. CF₃),optionally substituted O(C₁-C₆ alkyl) (preferably substituted with oneor two hydroxyl groups or up to three halo groups) or an optionallysubstituted acetylenic group —C≡C—R_(a) where R_(a) is H or a C₁-C₆alkyl group (preferably C₁-C₃ alkyl);R^(PRO) is H, optionally substituted C₁-C₆ alkyl or an optionallysubstituted aryl (phenyl or napthyl), heteroaryl or heterocyclic groupselected from the group consisting of oxazole, isoxazole, thiazole,isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine,furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,quinoline, (each preferably substituted with a C₁-C₃ alkyl group,preferably methyl or a halo group, preferably F or Cl), benzofuran,indole, indolizine, azaindolizine;R^(PRO1) and R^(PRO2) are each independently H, an optionallysubstituted C₁-C₃ alkyl group or together form a keto group, andEach n is independently 0, 1, 2, 3, 4, 5, or 6 (preferably 0 or 1);Each m′ is 0 or 1; andEach n′ is 0 or 1,Wherein each of said compounds, preferably on the alkyl, Aryl or Hetgroups, is substituted with a linker group to which is attached a

group (including a

group).

In alternative embodiments, R^(3′) is —(CH₂)_(n)-Aryl,—(CH₂CH₂O)_(n)-Aryl, —(CH₂)_(n)-HET or —(CH₂CH₂O)_(n)—HET;

Where Aryl is phenyl which is optionally substituted with one or twosubstitutents, wherein said substituent(s) is preferably selected from—(CH₂)_(n)OH, C₁-C₆ alkyl which itself is further optionally substitutedwith CN, halo (up to three halo groups), OH, —(CH₂)_(n)O(C₁-C₆)alkyl,amine, mono- or di-(C₁-C₆ alkyl) amine wherein the alkyl group on theamine is optionally substituted with 1 or 2 hydroxyl groups or up tothree halo (preferably F, Cl) groups, or said Aryl group is substitutedwith —(CH₂)_(n)OH, —(CH₂)_(n)—O—(C₁-C₆)alkyl,—(CH₂)_(n)—O—(CH₂)_(n)—(C₁-C₆)alkyl, —(CH₂)_(n)—C(O)(C₀-C₆) alkyl,—(CH₂)_(n)—C(O)O(C₀-C₆)alkyl, —(CH₂)_(n)—OC(O)(C₀-C₆)alkyl, amine, mono-or di-(C₁-C₆ alkyl) amine wherein the alkyl group on the amine isoptionally substituted with 1 or 2 hydroxyl groups or up to three halo(preferably F, Cl) groups, CN, NO₂, an optionally substituted—(CH₂)_(n)—(V)_(m′)—CH₂)_(n)—(V)_(m′)—(C₁-C₆)alkyl group, a—(V)_(m′)—(CH₂CH₂O)_(n)—R^(PEG) group where V is O, S or NR_(1′), R_(1′)is H or a C₁-C₃ alkyl group (preferably H) and R^(PEG) is H or a C₁-C₆alkyl group which is optionally substituted (including being optionallysubstituted with a carboxyl group), or said Aryl group is optionallysubstituted with a heterocycle, including a heteroaryl, selected fromthe group consisting of oxazole, isoxazole, thiazole, isothiazole,imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan,dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene,pyridine, piperidine, piperazine, morpholine, quinoline, benzofuran,indole, indolizine, azaindolizine, (when substituted each preferablysubstituted with a C₁-C₃ alkyl group, preferably methyl or a halo group,preferably F or Cl), or a group according to the chemical structure:

Where S^(c) is CHR^(SS), NR^(URE), or O;R^(HET) is H, CN, NO₂, halo (preferably Cl or F), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups (e.g. CF₃), optionally substituted O(C₁-C₆alkyl) (preferably substituted with one or two hydroxyl groups or up tothree halo groups) or an optionally substituted acetylenic group—C≡C—R_(a) where R_(a) is H or a C₁-C₆ alkyl group (preferably C₁-C₃alkyl);R^(SS) is H, CN, NO₂, halo (preferably F or Cl), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups), optionally substituted O—(C₁-C₆ alkyl)(preferably substituted with one or two hydroxyl groups or up to threehalo groups) or an optionally substituted —C(O)(C₁-C₆ alkyl) (preferablysubstituted with one or two hydroxyl groups or up to three halo groups);R^(URE) is H, a C₁-C₆ alkyl (preferably H or C₁-C₃ alkyl) or a—C(O)(C₀-C₆ alkyl), each of which groups is optionally substituted withone or two hydroxyl groups or up to three halogen, preferably fluorinegroups, or an optionally substituted heterocycle, for examplepiperidine, morpholine, pyrrolidine, tetrahydrofuran,tetrahydrothiophene, piperidine, piperazine, each of which is optionallysubstituted, andY^(C) is N or C—R^(YC), where R^(YC) is H, OH, CN, NO₂, halo (preferablyCl or F), optionally substituted C₁-C₆ alkyl (preferably substitutedwith one or two hydroxyl groups or up to three halo groups (e.g. CF₃),optionally substituted O(C₁-C₆ alkyl) (preferably substituted with oneor two hydroxyl groups or up to three halo groups) or an optionallysubstituted acetylenic group —C≡C—R_(a) where R_(a) is H or a C₁-C₆alkyl group (preferably C₁-C₃ alkyl);R^(PRO) is H, optionally substituted C₁-C₆ alkyl or an optionallysubstituted aryl (phenyl or napthyl), heteroaryl or heterocyclic groupselected from the group consisting of oxazole, isoxazole, thiazole,isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine,furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,quinoline, (each preferably substituted with a C₁-C₃ alkyl group,preferably methyl or a halo group, preferably F or Cl), benzofuran,indole, indolizine, azaindolizine;R^(PRO1) and R^(PRO2) are each independently H, an optionallysubstituted C₁-C₃ alkyl group or together form a keto group;HET is preferably oxazole, isoxazole, thiazole, isothiazole, imidazole,diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran,tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine,piperidine, piperazine, morpholine, quinoline, (each preferablysubstituted with a C₁-C₃ alkyl group, preferably methyl or a halo group,preferably F or Cl), benzofuran, indole, indolizine, azaindolizine, or agroup according to the chemical structure:

Where S^(c) is CHR^(SS), NR^(URE), or O;R^(HET) is H, CN, NO₂, halo (preferably Cl or F), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups (e.g. CF₃), optionally substituted O(C₁-C₆alkyl) (preferably substituted with one or two hydroxyl groups or up tothree halo groups) or an optionally substituted acetylenic group—C≡C—R_(a) where R_(a) is H or a C₁-C₆ alkyl group (preferably C₁-C₃alkyl);R^(SS) is H, CN, NO₂, halo (preferably F or Cl), optionally substitutedC₁-C₆ alkyl (preferably substituted with one or two hydroxyl groups orup to three halo groups), optionally substituted O—(C₁-C₆ alkyl)(preferably substituted with one or two hydroxyl groups or up to threehalo groups) or an optionally substituted —C(O)(C₁-C₆ alkyl) (preferablysubstituted with one or two hydroxyl groups or up to three halo groups);R^(URE) is H, a C₁-C₆ alkyl (preferably H or C₁-C₃ alkyl) or a—C(O)(C₀-C₆ alkyl), each of which groups is optionally substituted withone or two hydroxyl groups or up to three halogen, preferably fluorinegroups, or an optionally substituted heterocycle, for examplepiperidine, morpholine, pyrrolidine, tetrahydrofuran,tetrahydrothiophene, piperidine, piperazine, each of which is optionallysubstituted, andY^(C) is N or C—R^(YC), where R^(YC) is H, OH, CN, NO₂, halo (preferablyCl or F), optionally substituted C₁-C₆ alkyl (preferably substitutedwith one or two hydroxyl groups or up to three halo groups (e.g. CF₃),optionally substituted O(C₁-C₆ alkyl) (preferably substituted with oneor two hydroxyl groups or up to three halo groups) or an optionallysubstituted acetylenic group —C≡C—R_(a) where R_(a) is H or a C₁-C₆alkyl group (preferably C₁-C₃ alkyl);R^(PRO) is H, optionally substituted C₁-C₆ alkyl or an optionallysubstituted aryl, heteroaryl or heterocyclic group;R^(PRO1) and R^(PRO2) are each independently H, an optionallysubstituted C₁-C₃ alkyl group or together form a keto group,Each m′ is independently 0 or 1, andEach n is independently 0, 1, 2, 3, 4, 5, or 6 (preferably 0 or 1),Wherein each of said compounds, preferably on said Aryl or HET groups,is substituted with a linker group to which is attached a

group (including a

group).

In still additional embodiments, preferred compounds include thoseaccording to the chemical structure:

Where R^(1′) is OH or a group which is metabolized in a patient orsubject to OH;R^(2′) is a —NH—CH₂-Aryl-HET (preferably, a phenyl linked directly to amethyl substituted thiazole);R^(3′) is a —CHR^(CR3′)—NH—C(O)—R_(3P1) group or a —CHR^(CR3′)—R^(3P2)group;Where R^(CR3′) is a C₁-C₄ alkyl group, preferably methyl, isopropyl ortert-butyl;R^(3P1) is C₁-C₃ alkyl (preferably methyl), an optionally substitutedoxetane group (preferably methyl substituted, a —(CH₂)_(n)OCH₃ groupwhere n is 1 or 2 (preferably 2), or a

group (the ethyl ether group is preferably meta-substituted on thephenyl moiety), a morpholino grop (linked to the carbonyl at the 2- or3-position;

R^(3P2) is

group,Where Aryl is phenyl;HET is an optionally substituted thiazole or isothiazole; andR^(HET) is H or a halo group (preferably H),Or a pharmaceutically acceptable salt, stereoisomer, solvate orpolymorph thereof, wherein each of said compounds is substituted with alinker group to which is attached a

group (including a

group)

In an alternative embodiment,

groups for inclusion in compounds according to the present inventioninclude:

Where X is Cl, F, C₁-C₃ alkyl (preferably methyl) or heterocycle(preferably an optionally substituted heterocycle, including as definedabove for R^(3′);R¹ and R² are each independently H, C₁-C₃ alkyl (preferably methyl), orphenyl and each of said compounds is substituted with a linker group ora linker group to which is attached a

group, or a pharmaceutically acceptable salt, enantiomer, diasteromer,solvate or polymorph thereof.

Additional preferred

groups for inclusion in compounds according to the present inventioninclude:

Where n is 0 or 1;

R is a linker or a linker attached to a

groupX is H, F, Cl, C₁-C₃ alkyl (preferably methyl) or heterocycle(preferably an optionally substituted heterocycle, especially includinga water soluble heterocycle such as a morpholino group, including asdefined above for R^(3′)), ora pharmaceutically acceptable salt, enantiomer, diasteromer, solvate orpolymorph thereof.

Additional preferred

groups for inclusion in compounds according to the present inventioninclude for example:

Where n is 0 or 1;

R is a linker or a linker attached to a

group a linker or a linker attached to a

group linked to the

group through an amide, ester, ether or carbamate group;R¹ is C₁-C₃ alkyl (optionally substituted with one or two hydroxylgroups) or —C(O)NR³R₄ where R³ and R⁴ are each independently H, C₁-C₃alkyl (preferably methyl), phenyl or heterocycle (including aheterocycle such as a morpholino, piperazine or other group whichincreases water solubility),X is H, F, Cl, C₁-C₃ alkyl (preferably methyl) or heterocycle(preferably an optionally substituted heterocycle, including a watersoluble heterocycle, including as defined above for R_(3′)), or apharmaceutically acceptable salt, enantiomer, diasteromer, solvate orpolymorph thereof.

Still further preferred

groups for inclusion in compounds according to the present inventioninclude for example:

Where n is 0 or 1;

R¹ is a linker or a linker attached to a

group linked to the

group through an amide, ester, ether, carbamate or heterocyclic group(preferably an optionally substituted heterocycle, including as definedabove for R^(3′));R is H, F, Cl, C₁-C₃ alkyl (optionally substituted with one or twohydroxyl groups, preferably methyl), —O—C(O)NR³R⁴ or —C(O)NR³R⁴ whereineach of R³ and R⁴ is independently H, C₁-C₃ alkyl (preferably methyl),phenyl or heterocycle, including a water soluble heterocycle, or Apharmaceutically acceptable salt, enantiomer, diasteromer, solvate orpolymorph thereof.

Yet still further preferred

groups for inclusion in compounds according to the present inventioninclude for example:

Where n is 0 or 1;

R is a linker or a linker attached to a

group linked to the

group through an amide, ester, ether, carbamate or heterocyclic group;andEach X is independently is H, F, Cl, C₁-C₃ alkyl (optionally substitutedwith one or two hydroxyl groups, preferably methyl), heterocyle(preferably an optionally substituted heterocycle, including a watersoluble heterocycle, and/or as defined above for R^(3′)), —O—C(O)NR³R⁴or —C(O)NR³R⁴ wherein each of R³ and R⁴ is independently H, C₁-C₃ alkyl(preferably methyl), or phenyl, ora pharmaceutically acceptable salt, enantiomer, diasteromer, solvate orpolymorph thereof.

Yet additional further preferred

groups for inclusion in compounds according to the present inventioninclude for example:

Where n is 0 or 1;

R is a linker or a linker attached to a

group linked to the

group through an amide, ester, ether, carbamate or heterocyclic group;R¹ is C₁-C₃ alkyl, which is optionally substituted with one or twohydroxyl groups, —O—C(O)NR³R⁴ or —C(O)NR³R⁴ wherein each of R³ and R⁴ isindependently H, C₁-C₃ alkyl (preferably methyl), or phenyl; andX is independently is H, F, Cl, C₁-C₃ alkyl (optionally substituted withone or two hydroxyl groups, preferably methyl) or heterocycle(preferably an optionally substituted heterocycle, including a watersoluble heterocycle, and/or as defined above for R^(3′)), or apharmaceutically acceptable salt, enantiomer, diasteromer, solvate orpolymorph thereof.

Yet additional further preferred

groups for inclusion in compounds according to the present inventioninclude for example:

Where n is 0 or 1;

R is a linker or a linker attached to a

group linked to the

group through an amide, ester, ether, carbamate or heterocyclic group;R¹ is H, C₁-C₃ alkyl, which is optionally substituted with one or twohydroxyl groups, —O—C(O)NR³R⁴ or —C(O)NR³R⁴ wherein each of R³ and R⁴ isindependently H, C₁-C₃ alkyl (preferably methyl), or phenyl; andEach X is independently is H, F, Cl, C₁-C₃ alkyl (optionally substitutedwith one or two hydroxyl groups, preferably methyl) or heterocycle(preferably an optionally substituted heterocycle, including a watersoluble heterocycle and/or including as defined above for R^(3′)), or apharmaceutically acceptable salt, enantiomer, diastereomer, solvate orpolymorph thereof.

In an additional embodiment, particularly preferred compounds accordingto the present invention may be identified according to any one or moreof the chemical structures as set forth in FIG. 19 hereof:

Wherein any one or more of R_(1PC), R_(2PC), R_(3PC), R_(4PC), R_(5PC),R_(6PC), R_(7PC), R_(8PC), R_(9PC), R₁₀PC, R_(11PC), R_(12PC), R_(13PC)and R_(14PC) is a

group,Where L is a linker group and

is a protein targeting moiety, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthererof.

In preferred embodiments, no more than two of R_(1PC), R_(2PC), R_(3PC),R_(4PC), R_(5PC), R_(6PC), R_(7PC), R_(8PC), R_(9PC), R₁₀PC, R_(11PC),R_(12PC), R_(13PC) and R_(14PC) is a

group and the other of groups R_(1PC), R_(2PC), R_(3PC), R_(4PC),R_(5PC), R_(6PC), R_(7PC), R_(8PC), R_(9PC), R_(10PC), R_(11PC),R_(12PC), R_(13PC) and R_(14PC) are independently H or a CH₃ group,often H.

Certain preferred embodiments are directed to compounds according to thechemical structure:

Wherein R_(7PC) and R_(10PC) are each independently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, either of R_(7PC) Or R_(10PC) is a

group and the other R_(7PC) or R_(10PC) is H.

In other preferred embodiments, the compound has the chemical structure:

Wherein R_(7PC) is a

group, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC), R_(11PC) R_(12PC), R_(13PC) and R_(14PC) are eachindependently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC), R_(11PC), R_(12PC), R_(13PC) andR_(14PC) is a

group and the other groups are H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(4PC), R_(7PC), R_(11PC) R_(12PC), R_(13PC) and R_(14PC) areeach independently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, either of R_(4PC), R_(7PC) or one of R_(11PC),R_(12PC), R_(13PC) and R_(14PC) is a

group and the other groups are H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(3PC), R_(7PC) R_(11PC) R_(12PC), R_(13PC) and R_(14PC) areeach independently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(3PC), R_(7PC), R_(11PC), R_(12PC),R_(13PC) and R_(14PC) is a

group and the other groups are H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(10PC) are each independently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(10PC) is a

group and the other group is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(10PC) are each independently a

group or H and R_(8PC) is H or CH₃, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(10PC) is a

group and the other group is H and R_(8PC) is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(10PC) are each independently a

group or H and R_(8PC) is H or CH₃, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(10PC) is a

group and the other group is H and R_(8PC) is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(10PC) are each independently a

group or H and R_(8PC) is H or CH₃ orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(10PC) is a

group and the other group is H and R_(8PC) is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(10PC) are each independently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(10PC) is a

group and the other group is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(9PC) are each independently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(9PC) is a

group and the other group is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(14PC) are each independently a

group or H and each of R_(12PC) and R_(13PC) is H or CH₃, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(14PC) is a

group and the other of R_(7PC) and R_(14PC) group is H and each ofR_(12PC) and R_(13PC) is H,A pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(9PC) are each independently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(9PC) is a

group and the other group is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(9PC) are each independently a

group or H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably,one of R_(7PC) and R_(9PC) is a

group and the other group is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In still other preferred embodiments, the compound has the chemicalstructure:

Wherein R_(7PC) and R_(10PC) are each independently a

group or H and R_(9PC) is H or CH₃, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof. Preferably, one of R_(7PC) and R_(10PC) is a

group and the other group is H and R_(9PC) is H, orA pharmaceutically acceptable salt, stereoisomer, solvate or polymorphthereof.

In the above embodiments, the linker group may be any linker group asdescribed hereinabove, below is preferably a polyethylene glycol groupranging in size from about 1 to about 12 ethylene glycol units, between1 and about 10 ethylene glycol units, about 2 about 6 ethylene glycolunits, between about 2 and 5 ethylene glycol units, between about 2 and4 ethylene glycole units.

In preferred embodiments, the linker group L is a group:

Where Z is a group which link

to X; andX is a group linking Z to group

(including a

group).

In preferred aspects, Z is absent (a bond), —(CH₂)_(i)—O, —(CH₂)_(i)—S,—(CH₂)_(i)—N—R, a (CH₂)_(i)—X₁Y₁ group wherein X₁Y₁ forms an amidegroup, or a urethane group, ester or thioester group, or a

groupEach R is H, or a C₁-C₃ alkyl, an alkanol group or a heterocycle(including a water soluble heterocycle, preferably, a morpholino,piperidine or piperazine group to promote water solubility of the linkergroup);Each Y is independently a bond, O, S or N—R;and each i is independently 0 to 100, 1 to 75, 1 to 60, 1 to 55, 1 to50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to6, 1, 2, 3, 4 or 5;

In preferred aspects X is a

groupWhere each D is independently a bond absent),

j is 1 to 100, 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5;k is 1 to 100, 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5;preferably k is 1, 2, 3, 4, or 5;m′ is 1 to 100, 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5;n is 1 to 100, 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5;X¹ is O, S or N—R, preferably O;Y is the same as above; and

is a connector group (which may be a bond) which connects Z to X, whenpresent in the linker group.

In preferred aspects,

is a bond (absent), a heterocycle including a water soluble heterocyclesuch as a piperazinyl or other group or a

Where X² is O, S, NR⁴, S(O), S(O)₂, —S(O)₂O, —OS(O)₂, or OS(O)₂O;X³ is O, S, CHR⁴, NR⁴; andR⁴ is H or a C₁-C₃ alkyl group optionally substituted with one or twohydroxyl groups, or a pharmaceutically acceptable salt, enantiomer orstereoisomer thereof.

In alternative preferred aspects, the linker group is a(poly)ethyleneglycol having between 1 and about 100 ethylene glycolunits, between about 1 and about 50 ethylene glycol units, between 1 andabout 25 ethylene glycol units, between about 1 and 10 ethylene glycolunits, between 1 and about 8 ethylene glycol units and 1 and 6 ethyleneglycol units, between 2 and 4 ethylene glycol units.

In alternative preferred aspects,

is a

group or an amide group.

Although the

group and

group (including

group) may be covalently linked to the linker group through any groupwhich is appropriate and stable to the chemistry of the linker, inpreferred aspects of the present invention, the linker is independentlycovalently bonded to the

group and the

group (including a group) preferably through an amide, ester, thioester,keto group, carbamate

(urethane) or ether, each of which groups may be inserted anywhere onthe

group and

group (including a

group) to provide maximum binding of the

group on the ubiquitin ligase and the

group on the target protein to be degraded. (It is noted that in certainaspects where the

group is a

group, the target protein for degradation may be the ubiquitin ligaseitself). In certain preferred aspects, the linker may be linked to anoptionally substituted alkyl, alkylene, alkene or alkyne group, an arylgroup or a heterocyclic group on

the and/or

groups.

In preferred aspects of the invention, the

group is a

group, which binds to target proteins. Targets of the

group are numerous in kind and are selected from proteins that areexpressed in a cell such that at least a portion of the sequences isfound in the cell and may bind to a

group. The term “protein” includes oligopeptides and polypeptidesequences of sufficient length that they can bind to a

group according to the present invention. Any protein in a eukaryoticsystem or a microbial system, including a virus, bacteria or fungus, asotherwise described herein, are targets for ubiquitination mediated bythe compounds according to the present invention. Preferably, the targetprotein is a eukaryotic protein. In certain aspects, the protein bindingmoiety is a haloalkane (preferably a C1-C10 alkyl group which issubstituted with at least one halo group, preferably a halo group at thedistil end of the alkyl group (i.e., away from the linker or

group), which may covalently bind to a dehalogenase enzyme in a patientor subject or in a diagnostic assay.

groups according to the present invention include, for example, includeany moiety which binds to a protein specifically (binds to a targetprotein) and includes the following non-limiting examples of smallmolecule target protein moieties: Hsp90 inhibitors, kinase inhibitors,MDM2 inhibitors, compounds targeting Human BET Bromodomain-containingproteins, HDAC inhibitors, human lysine methyltransferase inhibitors,angiogenesis inhibitors, immunosuppressive compounds, and compoundstargeting the aryl hydrocarbon receptor (AHR), among numerous others.The compositions described below exemplify some of the members of thesenine types of small molecule target protein binding moieties. Such smallmolecule target protein binding moieties also include pharmaceuticallyacceptable salts, enantiomers, solvates and polymorphs of thesecompositions, as well as other small molecules that may target a proteinof interest. These binding moieties are linked to the ubiquitin ligasebinding moiety preferably through a linker in order to present a targetprotein (to which the protein target moiety is bound) in proximity tothe ubiquitin ligase for ubiquitination and degradation.

Any protein, which can bind to a protein target moiety or

group and acted on or degraded by an ubiquitin ligase is a targetprotein according to the present invention. In general, target proteinsmay include, for example, structural proteins, receptors, enzymes, cellsurface proteins, proteins pertinent to the integrated function of acell, including proteins involved in catalytic activity, aromataseactivity, motor activity, helicase activity, metabolic processes(anabolism and catrabolism), antioxidant activity, proteolysis,biosynthesis, proteins with kinase activity, oxidoreductase activity,transferase activity, hydrolase activity, lyase activity, isomeraseactivity, ligase activity, enzyme regulator activity, signal transduceractivity, structural molecule activity, binding activity (protein, lipidcarbohydrate), receptor activity, cell motility, membrane fusion, cellcommunication, regulation of biological processes, development, celldifferentiation, response to stimulus, behavioral proteins, celladhesion proteins, proteins involved in cell death, proteins involved intransport (including protein transporter activity, nuclear transport,ion transporter activity, channel transporter activity, carrieractivity, permease activity, secretion activity, electron transporteractivity, pathogenesis, chaperone regulator activity, nucleic acidbinding activity, transcription regulator activity, extracellularorganization and biogenesis activity, translation regulator activity.Proteins of interest can include proteins from eurkaryotes andprokaryotes including humans as targets for drug therapy, other animals,including domesticated animals, microbials for the determination oftargets for antibiotics and other antimicrobials and plants, and evenviruses, among numerous others.

In still other embodiments, the

group is a haloalkyl group, wherein said alkyl group generally ranges insize from about 1 or 2 carbons to about 12 carbons in length, oftenabout 2 to 10 carbons in length, often about 3 carbons to about 8carbons in length, more often about 4 carbons to about 6 carbons inlength. The haloalkyl groups are generally linear alkyl groups (althoughbranched-chain alkyl groups may also be used) and are end-capped with atleast one halogen group, preferably a single halogen group, often asingle chloride group. Haloalkyl

groups for use in the present invention are preferably represented bythe chemical structure —(CH₂)_(v)-Halo where v is any integer from 2 toabout 12, often about 3 to about 8, more often about 4 to about 6. Halomay be any halogen, but is preferably Cl or Br, more often Cl.

In still other embodiments, the

group is a

group, where w is 0 to 3, preferably 1 or 2. This group bindsselectively to estrogen receptors and is useful for treating diseaseswhich are modulated through estrogen receptors, and in particularcancers, such as breast cancer, endometrial cancer, ovarian cancer anduterine cancer, among others.

The present invention may be used to treat a number of disease statesand/or conditions, including any disease state and/or condition in whichproteins are dysregulated and where a patient would benefit from thedegradation of proteins.

In another aspect, the present invention relates to pharmaceuticalcompositions comprising an effective amount of a compound as set forthhereinabove, in combination with a pharmaceutically acceptable carrier,additive or excipient, and optionally an additional bioactive agent.

In alternative aspects, the present invention relates to a method fortreating a disease state by degrading a protein or polypeptide throughwhich a disease state or condition is modulated comprising administeringto said patient or subject an effective amount of at least one compoundas described hereinabove, optionally in combination with an additionalbioactive agent. The method according to the present invention may beused to treat a large number of disease states or conditions includingcancer, by virtue of the administration of effective amounts of at leastone compound described herein.

DETAILED DESCRIPTION OF THE INVENTION

The following terms are used to describe the present invention. Ininstances where a term is not specifically defined herein, that term isgiven an art-recognized meaning by those of ordinary skill applying thatterm in context to its use in describing the present invention.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise (such as in the case of a groupcontaining a number of carbon atoms in which case each carbon atomnumber falling within the range is provided), between the upper andlower limit of that range and any other stated or intervening value inthat stated range is encompassed within the invention. The upper andlower limits of these smaller ranges may independently be included inthe smaller ranges is also encompassed within the invention, subject toany specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either bothof those included limits are also included in the invention.

The term “compound”, as used herein, unless otherwise indicated, refersto any specific chemical compound disclosed herein and includestautomers, regioisomers, geometric isomers, and where applicable,stereoisomers, including optical isomers (enantiomers) and otherstereoisomers (diastereomers) thereof, as well as pharmaceuticallyacceptable salts and derivatives (including prodrug forms) thereof whereapplicable, in context. Within its use in context, the term compoundgenerally refers to a single compound, but also may include othercompounds such as stereoisomers, regioisomers and/or optical isomers(including racemic mixtures) as well as specific enantiomers orenantiomerically enriched mixtures of disclosed compounds. The term alsorefers, in context to prodrug forms of compounds which have beenmodified to facilitate the administration and delivery of compounds to asite of activity. It is noted that in describing the present compounds,numerous substituents and variables associated with same, among others,are described. It is understood by those of ordinary skill thatmolecules which are described herein are stable compounds as generallydescribed hereunder. When the bond

is shown, both a double bond and single bond are represented within thecontext of the compound shown.

The term “patient” or “subject” is used throughout the specification todescribe an animal, preferably a human or a domesticated animal, to whomtreatment, including prophylactic treatment, with the compositionsaccording to the present invention is provided.

For treatment of those infections, conditions or disease states whichare specific for a specific animal such as a human patient, the termpatient refers to that specific animal, including a domesticated animalsuch as a dog or cat or a farm animal such as a horse, cow, sheep, etc.In general, in the present invention, the term patient refers to a humanpatient unless otherwise stated or implied from the context of the useof the term.

The term “effective” is used to describe an amount of a compound,composition or component which, when used within the context of itsintended use, effects an intended result. The term effective subsumesall other effective amount or effective concentration terms, which areotherwise described or used in the present application.

The term “VCB E3 Ubiquitin Ligase”, “Hippel-Lindau E3 Ubiquitin Ligase”or “Ubiquitin Ligase” is used to describe a target enzyme(s) bindingsite of ubiquitin ligase moieties in the bifunctional (chimeric)compounds according to the present invention. VCB E3 is a protein thatin combination with an E2 ubiquitin-conjugating enzyme causes theattachment of ubiquitin to a lysine on a target protein; the E3ubiquitin ligase targets specific protein substrates for degradation bythe proteasome. Thus, E3 ubiquitin ligase alone or in complex with an E2ubiquitin conjugating enzyme is responsible for the transfer ofubiquitin to targeted proteins. In general, the ubiquitin ligase isinvolved in polyubiquitination such that a second ubiquitin is attachedto the first, a third is attached to the second, and so forth.Polyubiquitination marks proteins for degradation by the proteasome.However, there are some ubiquitination events that are limited tomono-ubiquitination, in which only a single ubiquitin is added by theubiquitin ligase to a substrate molecule. Mono-ubiquitinated proteinsare not targeted to the proteasome for degradation, but may instead bealtered in their cellular location or function, for example, via bindingother proteins that have domains capable of binding ubiquitin. Furthercomplicating matters, different lysines on ubiquitin can be targeted byan E3 to make chains. The most common lysine is Lys48 on the ubiquitinchain. This is the lysine used to make polyubiquitin, which isrecognized by the proteasome.

The term “protein target moiety” or PTM is used to describe a smallmolecule which binds to a target protein or other protein or polypeptideof interest and places/presents that protein or polypeptide in proximityto an ubiquitin ligase such that degradation of the protein orpolypeptide by ubiquitin ligase may occur. Non-limiting examples ofsmall molecule target protein binding moieties include Hsp90 inhibitors,kinase inhibitors, MDM2 inhibitors, compounds targeting Human BETBromodomain-containing proteins, HDAC inhibitors, human lysinemethyltransferase inhibitors, angiogenesis inhibitors, immunosuppressivecompounds, and compounds targeting the aryl hydrocarbon receptor (AHR),among numerous others. The compositions described below exemplify someof the members of these nine types of small molecule target protein.

Protein target moieties according to the present invention include, forexample, Haloalkane halogenase inhibitors, Hsp90 inhibitors, kinaseinhibitors, MDM2 inhibitors, compounds targeting Human BETBromodomain-containing proteins, HDAC inhibitors, human lysinemethyltransferase inhibitors, angiogenesis inhibitors, immunosuppressivecompounds, and compounds targeting the aryl hydrocarbon receptor (AHR).The compositions described below exemplify some of the members of thesetypes of small molecule target protein binding moieties. Such smallmolecule target protein binding moieties also include pharmaceuticallyacceptable salts, enantiomers, solvates and polymorphs of thesecompositions, as well as other small molecules that may target a proteinof interest. References which are cited hereinbelow are incorporated byreference herein in their entirety.

I. Heat Shock Protein 90 (HSP90) Inhibitors:

HSP90 inhibitors as used herein include, but are not limited to:1. The HSP90 inhibitors identified in Vallee, et al., “Tricyclic Seriesof Heat Shock Protein 90 (HSP90) Inhibitors Part I: Discovery ofTricyclic Imidazo[4,5-C]Pyridines as Potent Inhibitors of the HSP90Molecular Chaperone (2011) J. Med. Chem. 54: 7206, including

Derivatized where a linker group L or a

group is attached via the terminal amide group;

2. The HSP90 Inhibitor p54 (Modified):

Where a linker group L or a

group is attached via the terminal acetylene group;3. The HSP90 inhibitors (modified) identified in Brough, et al.,“4,5-Diarylisoxazole HSP90 Chaperone Inhibitors: Potential TherapeuticAgents for the Treatment of Cancer”, J. MED. CHEM. vol: 51, page: 196(2008), including the compound 2GJ(5-[2,4-DIHYDROXY-5-(1-METHYLETHYL)PHENYL]-N-ETHYL-4-[4-(MORPHOLIN-4-YLMETHYL)PHENYL]ISOXAZOLE-3-CARBOXAMIDE)having the structure:

Derivatized, where a linker group L or a

group is attached via the amide group (at the amine or at the alkylgroup on the amine;4. The HSP90 inhibitors (modified) identified in Wright, et al.,Structure-Activity Relationships in Purine-Based Inhibitor Binding toHSP90 Isoforms, Chem Biol. 2004 June; 11(6):775-85, including the HSP90inhibitor PU3 having the structure:

Where a linker group L or

is attached via the butyl group; and5. The HSP90 inhibitor Geldanamycin((4E,6Z,8S,9S,10E,12S,13R,14S,16R)-13-hydroxy-8,14,19-trimethoxy-4,10,12,16-tetramethyl-3,20,22-trioxo-2-azabicyclo[16.3.1](derivatized)or any of its derivatives (e.g. 17-alkylamino-17-desmethoxygeldanamycin(“17-AAG”) or 17-(2-dimethylaminoethyl)amino-17-desmethoxygeldanamycin(“17-DMAG”)) (derivatized, where a linker group L or a

group is attached via the amide group).

II. Kinase and Phosphatase Inhibitors:

Kinase inhibitors as used herein include, but are not limited to:

1. Erlotinib Derivative Tyrosine Kinase Inhibitor

Where R is a linker group L or a

group attached via the ether group;2. The kinase inhibitor Sunitanib (derivatized):

(Derivatized where R is a linker group L or a

group attached to the pyrrole moiety);13. Kinase inhibitor Sorafenib (derivatized)

(Derivatized where R is a linker group L or a

group attached to the phenyl moiety);4. The kinase Inhibitor Desatinib (derivatized)

(Derivatized where R is a linker group L or a

attached to the pyrimidine);5. The kinase inhibitor Lapatinib (derivatized)

Derivatized where a linker group L or a

group is attached via the terminal methyl of the sulfonyl methyl group;6. The kinase inhibitor U09-CX-5279 (Derivatized)7.

Derivatized where a linker group L or a

group is attached via the amine (aniline), carboxylic acid or aminealpha to cyclopropyl group, or cyclopropyl group;7. The kinase inhibitors identified in Millan, et al., Design andSynthesis of Inhaled P38 Inhibitors for the Treatment of ChronicObstructive Pulmonary Disease, J. MED. CHEM. vol: 54, page: 7797 (2011),including the kinase inhibitors Y1W and Y1X (Derivatized) having thestructures:

Derivatized where a linker group L or a

group is attached preferably via the propyl group;

Derivatized where a linker group L or a

group is attached preferably via either the propyl group or the butylgroup;8. The kinase inhibitors identified in Schenkel, et al., Discovery ofPotent and Highly Selective Thienopyridine Janus Kinase 2 Inhibitors J.Med. Chem., 2011, 54 (24), pp 8440-8450, including the compounds 6TP and0TP (Derivatized) having the structures:

Derivatized where a linker group L or a

group is attached preferably via the terminal methyl group bound toamide moiety;

Derivatized where a linker group L or a

group is attached preferably via the terminal methyl group bound to theamide moiety;9. The kinase inhibitors identified in Van Eis, et al.,“2,6-Naphthyridines as potent and selective inhibitors of the novelprotein kinase C isozymes”, Biorg. Med. Chem. Lett. 2011 Dec. 15;21(24):7367-72, including the kinase inhibitor 07U having the structure:

Derivatized where a linker group L or a

group is attached preferably via the secondary amine or terminal aminogroup;10. The kinase inhibitors identified in Lountos, et al., “StructuralCharacterization of Inhibitor Complexes with Checkpoint Kinase 2 (Chk2),a Drug Target for Cancer Therapy”, J. STRUCT. BIOL. vol: 176, page: 292(2011), including the kinase inhibitor YCF having the structure:

Derivatized where a linker group L or a

group is attached preferably via either of the terminal hydroxyl groups;11. The kinase inhibitors identified in Lountos, et al., “StructuralCharacterization of Inhibitor Complexes with Checkpoint Kinase 2 (Chk2),a Drug Target for Cancer Therapy”, J. STRUCT. BIOL. vol: 176, page: 292(2011), including the kinase inhibitors XK9 and NXP (derivatized) havingthe structures:

Derivatized where a linker group L or a

group is attached preferably via the terminal hydroxyl group (XK9) orthe hydrazone group (NXP);12. The kinase inhibitor Afatinib (derivatized)(N-[4-[(3-Chloro-4-fluorophenyl)amino]-7-[[(3S)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4(dimethylamino)-2-butenamide)(Derivatized where a linker group L or a

group is attached preferably via the aliphatic amine group);13. The kinase inhibitor Fostamatinib (derivatized)([6-({5-fluoro-2-[(3,4,5-trimethoxyphenyl)amino]pyrimidin-4-yl}amino)-2,2-dimethyl-3-oxo-2,3-dihydro-4H-pyrido[3,2-b]-1,4-oxazin-4-yl]methyldisodium phosphate hexahydrate) (Derivatized where a linker group L or a

group is attached preferably via a methoxy group);14. The kinase inhibitor Gefitinib (derivatized)(N-(3-chloro-4-fluoro-phenyl)-7-methoxy-6-(3-morpholin-4-ylpropoxy)quinazolin-4-amine)(Derivatized where a linker group L or a

group is attached preferably via a methoxy or ether group);

15. The kinase inhibitor Lenvatinib (derivatized)(4-[3-chloro-4-(cyclopropylcarbamoylamino)phenoxy]-7-methoxy-quinoline-6-carboxamide)(Derivatized where a linker group L or a

group is attached preferably via the cyclopropyl group);16. The kinase inhibitor Vandetanib (derivatized)(N-(4-bromo-2-fluorophenyl)-6-methoxy-7-[(1-methylpiperidin-4-yl)methoxy]quinazolin-4-amine)(derivatized where a linker group L or a

group is attached preferably via the methoxy or hydroxyl group); and17. The kinase inhibitor Vemurafenib (derivatized) (propane-1-sulfonicacid{3-[5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl]-2,4-difluoro-phenyl}-amide)(Derivatized where a linker group L or a

group is attached preferably via the sulfonyl propyl group);18. The kinase inhibitor Gleevec (derivatized):

(Derivatized where R as a linker group L or a

group is attached preferably via the amide group or via the anilineamine group);19. The kinase inhibitor Pazopanib (derivatized) (VEGFR3 inhibitor):

(Derivatized where R is a linker group L or a

group preferably attached to the phenyl moiety or via the aniline aminegroup);20. The kinase inhibitor AT-9283 (Derivatized) Aurora Kinase Inhibitor

(where R is a linker group L or a

group attached preferably to the phenyl moiety);21. The kinase inhibitor TAE684 (derivatized) ALK inhibitor

(where R is a linker group L or a

group attached preferably to the phenyl moiety);22. The kinase inhibitor Nilotanib (derivatized) Abl inhibitor:

(Derivatized where R is a linker group L or a

group attached preferably to the phenyl moiety or the aniline aminegroup);27. Kinase Inhibitor NVP-BSK805 (derivatized) JAK2 Inhibitor

(Derivatized where R is a linker group L or a

group attached to the phenyl moiety or the diazole group);

28. Kinase Inhibitor Crizotinib Derivatized Alk Inhibitor

(Derivatized where R is a linker group L or a

group attached to the phenyl moiety or the diazole group);29. Kinase Inhibitor JNJ FMS (derivatized) Inhibitor

(Derivatized where R is a linker group L or a

group attached preferably to the phenyl moiety);30. The kinase inhibitor Foretinib (derivatized) Met Inhibitor

(Derivatized where R is a linker group L or a

group attached to the phenyl moiety or a hydroxyl or ether group on thequinoline moiety);31. The allosteric Protein Tyrosine Phosphatase Inhibitor PTP1B(derivatized):

Derivatized where a linker group L or a

group is preferably attached at R, as indicated.32. The inhibitor of SHP-2 Domain of Tyrosine Phosphatase (derivatized):

Derivatized where a linker group L or a

group is attached preferably at R.33. The inhibitor (derivatized) of BRAF (BRAF^(V600E))/MEK:

Derivatized where a linker group L or a

group is attached preferably at R.34. Inhibitor (derivatized) of Tyrosine Kinase ABL

(Derivatized where “R” designates a site for attachment of a linkergroup L or a

group on the piperazine moiety).

III. MDM2 Inhibitors:

MDM2 inhibitors as used herein include, but are not limited to:1. The MDM2 inhibitors identified in Vassilev, et al., In vivoactivation of the p53 pathway by small-molecule antagonists of MDM2,SCIENCE vol: 303, page: 844-848 (2004), and Schneekloth, et al.,Targeted intracellular protein degradation induced by a small molecule:En route to chemical proteomics, Bioorg. Med. Chem. Lett. 18 (2008)5904-5908, including (or additionally) the compounds nutlin-3, nutlin-2,and nutlin-1 (derivatized) as described below, as well as allderivatives and analogs thereof:

(Derivatized where a linker group L or a

group is attached preferably at the methoxy group or as a hydroxylgroup)

(Derivatized where a linker group L or a

group is attached preferably at the methoxy group or hydroxyl group);

(Derivatized where a linker group L or a

group is attached via the methoxy group or as a hydroxyl group); and

2. Trans-4-Iodo-4′-Boranyl-Chalcone

(Derivatized where a linker group L or a

group is attached a linker group L or a

group is attached group is attached via a hydroxy group);

IV. Compounds Targeting Human BET Bromodomain-Containing Proteins:

Compounds targeting Human BET Bromodomain-containing proteins include,but are not limited to the compounds associated with the targets asdescribed below, where “R” designates a site for linker group L or a

group attachment: 1.

Protein Targets: Brd2, Brd3, Brd4

JQ1, Filippakopoulos et al. Selective inhibition of BET bromodomains.Nature (2010)2.

I-BET, Nicodeme et al. Suppression of inflammation by a synthetichistone mimic. Nature (2010) Chung et al. Discovery and Characterizationof Small Molecule Inhibitors of the BET Family Bromodomains. Journal ofmedicinal chemistry (2011)3.

4d, Hewings et al. 3,5-Dimethylisoxazoles Act As Acetyl-lysine-mimeticBromodomain Ligands. J. Med. Chem. (2011) vol. 54 (19) pp. 6761-704.

I-BET151, Dawson et al. Inhibition of BET recruitment to chromatin as aneffective treatment for MLL-fusion leukaemia. Nature (2011)Where R, in each instance, designates a site for attachment of a linkergroup L or a

group).

V. HDAC Inhibitors:

HDAC Inhibitors (derivatized) include, but are not limited to:

1.

Finnin, M. S. et al. Structures of a histone deacetylase homologue boundto the TSA and SAHA inhibitors. Nature 401, 188-193 (1999).(Derivatized where “R” designates a site for attachment of a linkergroup L or a

group); and2. Compounds as defined by formula (I) of PCT WO0222577 (“DEACETYLASEINHIBITORS”) (Derivatized where a linker group L or a

group is attached via the hydroxyl group);

VI. Human Lysine Methyltransferase Inhibitors:

Human Lysine Methyltransferase inhibitors include, but are not limitedto:

1.

Chang et al. Structural basis for G9a-like protein lysinemethyltransferase inhibition by BIX-01294. Nat Struct Mol Biol (2009)vol. 16 (3) pp. 312-7(Derivatized where “R” designates a site for attachment of a linkergroup L or a

group);

Liu F, Chen X, Allali-Hassani A, et al. Discovery of a2,4-diamino-7-aminoalkoxyquinazoline as a potent and selective inhibitorof histone lysine methyltransferase G9a. J Med Chem 2009; 52(24):7950-3(Derivatized where “R” designates a potential site for attachment of alinker group L or a

group);3. Azacitidine (derivatized)(4-amino-1-β-D-ribofuranosyl-1,3,5-triazin-2(1H)-one) (Derivatized wherea linker group L or a

group is attached via the hydroxy or amino groups); and4. Decitabine (derivatized)(4-amino-1-(2-deoxy-b-D-erythro-pentofuranosyl)-1,3,5-triazin-2(1H)-one)(Derivatized where a linker group L or a

group is attached via either of the hydroxy groups or at the aminogroup).

VII. Angiogenesis Inhibitors:

Angiogenesis inhibitors include, but are not limited to:1. GA-1 (derivatized) and derivatives and analogs thereof, having thestructure(s) and binding to linkers as described in Sakamoto, et al.,Development of Protacs to target cancer-promoting proteins forubiquitination and degradation, Mol Cell Proteomics 2003 December;2(12):1350-8;2. Estradiol (derivatized), which may be bound to a linker group L or a

group as is generally described in Rodriguez-Gonzalez, et al., Targetingsteroid hormone receptors for ubiquitination and degradation in breastand prostate cancer, Oncogene (2008) 27, 7201-7211;3. Estradiol, testosterone (derivatized) and related derivatives,including but not limited to DHT and derivatives and analogs thereof,having the structure(s) and binding to a linker group L or a

group as generally described in Sakamoto, et al., Development of Protacsto target cancer-promoting proteins for ubiquitination and degradation,Mol Cell Proteomics 2003 December; 2(12): 1350-8; and4. Ovalicin, fumagillin (derivatized), and derivatives and analogsthereof, having the structure(s) and binding to a linker group L or a

group as is generally described in Sakamoto, et al., Protacs: chimericmolecules that target proteins to the Skp1-Cullin-F box complex forubiquitination and degradation Proc Natl Acad Sci USA. 2001 Jul. 17;98(15):8554-9 and U.S. Pat. No. 7,208,157.

VIII. Immunosuppressive Compounds:

Immunosuppressive compounds include, but are not limited to:1. AP21998 (derivatized), having the structure(s) and binding to alinker group L or a

group as is generally described in Schneekloth, et al., Chemical GeneticControl of Protein Levels: Selective in Vivo Targeted Degradation, J.AM. CHEM. SOC. 2004, 126, 3748-3754;2. Glucocorticoids (e.g., hydrocortisone, prednisone, prednisolone, andmethylprednisolone) (Derivatized where a linker group L or a

group is to bound, e.g. to any of the hydroxyls) and beclometasonedipropionate (Derivatized where a linker group or a

is bound, e.g. to a proprionate);3. Methotrexate (Derivatized where a linker group or a

group can be bound, e.g. to either of the terminal hydroxyls);4. Ciclosporin (Derivatized where a linker group or a

group can be bound, e.g. at any of the butyl groups);5. Tacrolimus (FK-506) and rapamycin (Derivatized where a linker group Lor a

group can be bound, e.g. at one of the methoxy groups); and6. Actinomycins (Derivatized where a linker group L or a

group can be bound, e.g. at one of the isopropyl groups).

IX. Compounds Targeting the Aryl Hydrocarbon Receptor (AHR):

Compounds targeting the aryl hydrocarbon receptor (AHR) include, but arenot limited to:

1. Apigenin (Derivatized in a way which binds to a linker group L or a

group as is generally illustrated in Lee, et al., Targeted Degradationof the Aryl Hydrocarbon Receptor by the PROTAC Approach: A UsefulChemical Genetic Tool, ChemBioChem Volume 8, Issue 17, pages 2058-2062,Nov. 23, 2007); and2. SR1 and LGC006 (derivatized such that a linker group L or a

is bound), as described in Boitano, et al., Aryl Hydrocarbon ReceptorAntagonists Promote the Expansion of Human Hematopoietic Stem Cells,Science 10 Sep. 2010: Vol. 329 no. 5997 pp. 1345-1348.

X. Compounds Targeting RAF Receptor (Kinase):

(Derivatized where “R” designates a site for linker group L or

group attachment).

XI. Compounds Targeting FKBP

(Derivatized where “R” designates a site for a linker group L or a

group attachment).

XII. Compounds Targeting Androgen Receptor (AR)

1. RU59063 Ligand (derivatized) of Androgen Receptor

(Derivatized where “R” designates a site for a linker group L or a

group attachment).2. SARM Ligand (derivatized) of Androgen Receptor

(Derivatized where “R” designates a site for a linker group L or a

group attachment).3. Androgen Receptor Ligand DHT (derivatized)

(Derivatized where “R” designates a site for a linker group L or

group attachment).

XIII. Compounds Targeting Estrogen Receptor (ER) ICI-182780 1. EstrogenReceptor Ligand

(Derivatized where “R” designates a site for linker group L or

group attachment).

XIV. Compounds Targeting Thyroid Hormone Receptor (TR) 1. ThyroidHormone Receptor Ligand (Derivatized)

(Derivatized where “R” designates a site for linker group L or

group attachment and MOMO indicates a methoxymethoxy group).XV. Compounds targeting HIV Protease

1. Inhibitor of HIV Protease (Derivatized)

(Derivatized where “R” designates a site for linker group L or

group attachment). See, J. Med. Chem. 2010, 53, 521-538.

2. Inhibitor of HIV Protease

(Derivatized where “R” designates a potential site for linker group L or

group attachment). See, J. Med. Chem. 2010, 53, 521-538.

XVI. Compounds Targeting HIV Integrase 1. Inhibitor of HIV Integrase(Derivatized)

(Derivatized where “R” designates a site for linker group L or

group attachment). See, J. Med. Chem. 2010, 53, 6466.

2. Inhibitor of HIV Integrase (Derivatized)

(Derivatized where “R” designates a site for linker group L or

group attachment). See, J. Med. Chem. 2010, 53, 6466.XVII. Compounds targeting HCV Protease

1. Inhibitors of HCV Protease (Derivatized)

(Derivatized where “R” designates a site for linker group L or

group attachment).

XVIII. Compounds Targeting Acyl-Protein Thioesterase-1 and -2 (APT1 andAPT2) 1. Inhibitor of APT1 and APT2 (Derivatized)

(Derivatized where “R” designates a site for linker group L or

group attachment). See, Angew. Chem. Int. Ed. 2011, 50, 9838-9842, whereL is a linker group as otherwise described herein and said

group is as otherwise described herein such that

binds the

group to a

group as otherwise described herein.

The term “target protein” is used to describe a protein or polypeptide,which is a target for binding to a compound according to the presentinvention and degradation by ubiquitin ligase hereunder. Such smallmolecule target protein binding moieties also include pharmaceuticallyacceptable salts, enantiomers, solvates and polymorphs of thesecompositions, as well as other small molecules that may target a proteinof interest. These binding moieties are linked to

groups through linker groups L.

Target proteins which may be bound to the protein target moiety anddegraded by the ligase to which the ubiquitin ligase binding moiety isbound include structural proteins, receptors, enzymes, cell surfaceproteins, proteins pertinent to the integrated function of a cell,including proteins involved in catalytic activity, aromatase activity,motor activity, helicase activity, metabolic processes (anabolism andcatrabolism), antioxidant activity, proteolysis, biosynthesis, proteinswith kinase activity, oxidoreductase activity, transferase activity,hydrolase activity, lyase activity, isomerase activity, ligase activity,enzyme regulator activity, signal transducer activity, structuralmolecule activity, binding activity (protein, lipid carbohydrate),receptor activity, cell motility, membrane fusion, cell communication,regulation of biological processes, development, cell differentiation,response to stimulus, behavioral proteins, cell adhesion proteins,proteins involved in cell death, proteins involved in transport(including protein transporter activity, nuclear transport, iontransporter activity, channel transporter activity, carrier activity,permease activity, secretion activity, electron transporter activity,pathogenesis, chaperone regulator activity, nucleic acid bindingactivity, transcription regulator activity, extracellular organizationand biogenesis activity, translation regulator activity. Proteins ofinterest can include proteins from eurkaryotes and prokaryotes,including microbes, viruses, fungi and parasites, including humans,microbes, viruses, fungi and parasites, among numerous others, astargets for drug therapy, other animals, including domesticated animals,microbials for the determination of targets for antibiotics and otherantimicrobials and plants, and even viruses, among numerous others

More specifically, a number of drug targets for human therapeuticsrepresent protein targets to which protein target moiety may be boundand incorporated into compounds according to the present invention.These include proteins which may be used to restore function in numerouspolygenic diseases, including for example B7.1 and B7, TINFRlm, TNFR2,NADPH oxidase, BclIBax and other partners in the apotosis pathway, C5areceptor, HMG-CoA reductase, PDE V phosphodiesterase type, PDE IVphosphodiesterase type 4, PDE I, PDEII, PDEIII, squalene cyclaseinhibitor, CXCR1, CXCR2, nitric oxide (NO) synthase, cyclo-oxygenase 1,cyclo-oxygenase 2, 5HT receptors, dopamine receptors, G Proteins, i.e.,Gq, histamine receptors, 5-lipoxygenase, tryptase serine protease,thymidylate synthase, purine nucleoside phosphorylase, GAPDHtrypanosomal, glycogen phosphorylase, Carbonic anhydrase, chemokinereceptors, JAW STAT, RXR and similar, HIV 1 protease, HIV 1 integrase,influenza, neuramimidase, hepatitis B reverse transcriptase, sodiumchannel, multi drug resistance (MDR), protein P-glycoprotein (and MRP),tyrosine kinases, CD23, CD124, tyrosine kinase p56 lck, CD4, CD5, IL-2receptor, IL-1 receptor, TNF-alphaR, ICAM1, Cat+ channels, VCAM, VLA-4integrin, selectins, CD40/CD40L, newokinins and receptors, inosinemonophosphate dehydrogenase, p38 MAP Kinase, RaslRaflMEWERK pathway,interleukin-1 converting enzyme, caspase, HCV, NS3 protease, HCV NS3 RNAhelicase, glycinamide ribonucleotide formyl transferase, rhinovirus 3Cprotease, herpes simplex virus-1 (HSV-I), protease, cytomegalovirus(CMV) protease, poly (ADP-ribose) polymerase, cyclin dependent kinases,vascular endothelial growth factor, oxytocin receptor, microsomaltransfer protein inhibitor, bile acid transport inhibitor, 5 alphareductase inhibitors, angiotensin 11, glycine receptor, noradrenalinereuptake receptor, endothelin receptors, neuropeptide Y and receptor,estrogen receptors, androgen receptors, adenosine receptors, adenosinekinase and AMP deaminase, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6,P2X1-7), farnesyltransferases, geranylgeranyl transferase, TrkA areceptor for NGF, beta-amyloid, tyrosine kinase Flk-IIKDR, vitronectinreceptor, integrin receptor, Her-21 neu, telomerase inhibition,cytosolic phospholipaseA2 and EGF receptor tyrosine kinase. Additionalprotein targets include, for example, ecdysone 20-monooxygenase, ionchannel of the GABA gated chloride channel, acetylcholinesterase,voltage-sensitive sodium channel protein, calcium release channel, andchloride channels. Still further target proteins include Acetyl-CoAcarboxylase, adenylosuccinate synthetase, protoporphyrinogen oxidase,and enolpyruvylshikimate-phosphate synthase.

Haloalkane dehalogenase enzymes are another target of specific compoundsaccording to the present invention. Compounds according to the presentinvention which contain chloroalkane peptide binding moieties (C₁-C₁₂often about C₂-C₁₀ alkyl halo groups) may be used to inhibit and/ordegrade haloalkane dehalogenase enzymes which are used in fusionproteins or related dioagnostic proteins as described in PCT/US2012/063401 filed Dec. 6, 2011 and published as WO 2012/078559 on Jun.14, 2012, the contents of which is incorporated by reference herein.

These various protein targets may be used in screens that identifycompound moieties which bind to the protein and by incorporation of themoiety into compounds according to the present invention, the level ofactivity of the protein may be altered for therapeutic end result.

The term “disease state or condition” is used to describe any diseasestate or condition wherein protein dysregulation (i.e., the amount ofprotein expressed in a patient is elevated) occurs and where degradationof one or more proteins in a patient may provide beneficial therapy orrelief of symptoms to a patient in need thereof. In certain instances,the disease state or condition may be cured.

Disease states of conditions which may be treated using compoundsaccording to the present invention include, for example, asthma,autoimmune diseases such as multiple sclerosis, various cancers,ciliopathies, cleft palate, diabetes, heart disease, hypertension,inflammatory bowel disease, mental retardation, mood disorder, obesity,refractive error, infertility, Angelman syndrome, Canavan disease,Coeliac disease, Charcot-Marie-Tooth disease, Cystic fibrosis, Duchennemuscular dystrophy, Haemochromatosis, Haemophilia, Klinefelter'ssyndrome, Neurofibromatosis, Phenylketonuria, Polycystic kidney disease,(PKD1) or 4 (PKD2) Prader-Willi syndrome, Sickle-cell disease, Tay-Sachsdisease, Turner syndrome.

Further disease states or conditions which may be treated by compoundsaccording to the present invention include Alzheimer's disease,Amyotrophic lateral sclerosis (Lou Gehrig's disease), Anorexia nervosa,Anxiety disorder, Atherosclerosis, Attention deficit hyperactivitydisorder, Autism, Bipolar disorder, Chronic fatigue syndrome, Chronicobstructive pulmonary disease, Crohn's disease, Coronary heart disease,Dementia, Depression, Diabetes mellitus type 1, Diabetes mellitus type2, Epilepsy, Guillain-Barre syndrome, Irritable bowel syndrome, Lupus,Metabolic syndrome, Multiple sclerosis, Myocardial infarction, Obesity,Obsessive-compulsive disorder, Panic disorder, Parkinson's disease,Psoriasis, Rheumatoid arthritis, Sarcoidosis, Schizophrenia, Stroke,Thromboangiitis obliterans, Tourette syndrome, Vasculitis.

Still additional disease states or conditions which can be treated bycompounds according to the present invention include aceruloplasminemia,Achondrogenesis type II, achondroplasia, Acrocephaly, Gaucher diseasetype 2, acute intermittent porphyria, Canavan disease, AdenomatousPolyposis Coli, ALA dehydratase deficiency, adenylosuccinate lyasedeficiency, Adrenogenital syndrome, Adrenoleukodystrophy, ALA-Dporphyria, ALA dehydratase deficiency, Alkaptonuria, Alexander disease,Alkaptonuric ochronosis, alpha 1-antitrypsin deficiency, alpha-1proteinase inhibitor, emphysema, amyotrophic lateral sclerosis Alstromsyndrome, Alexander disease, Amelogenesis imperfecta, ALA dehydratasedeficiency, Anderson-Fabry disease, androgen insensitivity syndrome,Anemia Angiokeratoma Corporis Diffusum, Angiomatosis retinae (vonHippel-Lindau disease) Apert syndrome, Arachnodactyly (Marfan syndrome),Stickler syndrome, Arthrochalasis multiplex congenital (Ehlers-Danlossyndrome#arthrochalasia type) ataxia telangiectasia, Rett syndrome,primary pulmonary hypertension, Sandhoff disease, neurofibromatosis typeII, Beare-Stevenson cutis gyrata syndrome, Mediterranean fever,familial, Benjamin syndrome, beta-thalassemia Bilateral AcousticNeurofibromatosis (neurofibromatosis type II), factor V Leidenthrombophilia, Bloch-Sulzberger syndrome (incontinentia pigmenti), Bloomsyndrome, X-linked sideroblastic anemia, Bonnevie-Ullrich syndrome(Turner syndrome), Bourneville disease (tuberous sclerosis), priondisease, Birt-Hogg-Dube syndrome, Brittle bone disease (osteogenesisimperfecta), Broad Thumb-Hallux syndrome (Rubinstein-Taybi syndrome),Bronze Diabetes/Bronzed Cirrhosis (hemochromatosis), Bulbospinalmuscular atrophy (Kennedy's disease), Burger-Grutz syndrome (lipoproteinlipase deficiency), CGD Chronic granulomatous disorder, Campomelicdysplasia, biotinidase deficiency, Cardiomyopathy (Noonan syndrome), Cridu chat, CAVD (congenital absence of the vas deferens), Caylorcardiofacial syndrome (CBAVD), CEP (congenital erythropoieticporphyria), cystic fibrosis, congenital hypothyroidism, Chondrodystrophysyndrome (achondroplasia), otospondylomegaepiphyseal dysplasia,Lesch-Nyhan syndrome, galactosemia, Ehlers-Danlos syndrome,Thanatophoric dysplasia, Coffin-Lowry syndrome, Cockayne syndrome,(familial adenomatous polyposis), Congenital erythropoietic porphyria,Congenital heart disease, Methemoglobinemia/Congenitalmethaemoglobinaemia, achondroplasia, X-linked sideroblastic anemia,Connective tissue disease, Conotruncal anomaly face syndrome, Cooley'sAnemia (beta-thalassemia), Copper storage disease (Wilson's disease),Copper transport disease (Menkes disease), hereditary coproporphyria,Cowden syndrome, Craniofacial dysarthrosis (Crouzon syndrome),Creutzfeldt-Jakob disease (prion disease), Cockayne syndrome, Cowdensyndrome, Curschmann-Batten-Steinert syndrome (myotonic dystrophy),Beare-Stevenson cutis gyrata syndrome, primary hyperoxaluria,spondyloepimetaphyseal dysplasia (Strudwick type), muscular dystrophy,Duchenne and Becker types (DBMD), Usher syndrome, Degenerative nervediseases including de Grouchy syndrome and Dejerine-Sottas syndrome,developmental disabilities, distal spinal muscular atrophy, type V,androgen insensitivity syndrome, Diffuse Globoid Body Sclerosis (Krabbedisease), Di George's syndrome, Dihydrotestosterone receptor deficiency,androgen insensitivity syndrome, Down syndrome, Dwarfism, erythropoieticprotoporphyria Erythroid 5-aminolevulinate synthetase deficiency,Erythropoietic porphyria, erythropoietic protoporphyria, erythropoieticuroporphyria, Friedreich's ataxia, familial paroxysmal polyserositis,porphyria cutanea tarda, familial pressure sensitive neuropathy, primarypulmonary hypertension (PPH), Fibrocystic disease of the pancreas,fragile X syndrome, galactosemia, genetic brain disorders, Giant cellhepatitis (Neonatal hemochromatosis), Gronblad-Strandberg syndrome(pseudoxanthoma elasticum), Gunther disease (congenital erythropoieticporphyria), haemochromatosis, Hallgren syndrome, sickle cell anemia,hemophilia, hepatoerythropoietic porphyria (HEP), Hippel-Lindau disease(von Hippel-Lindau disease), Huntington's disease, Hutchinson-Gilfordprogeria syndrome (progeria), Hyperandrogenism, Hypochondroplasia,Hypochromic anemia, Immune system disorders, including X-linked severecombined immunodeficiency, Insley-Astley syndrome, Jackson-Weisssyndrome, Joubert syndrome, Lesch-Nyhan syndrome, Jackson-Weisssyndrome, Kidney diseases, including hyperoxaluria, Klinefelter'ssyndrome, Kniest dysplasia, Lacunar dementia, Langer-Saldinoachondrogenesis, ataxia telangiectasia, Lynch syndrome,Lysyl-hydroxylase deficiency, Machado-Joseph disease, Metabolicdisorders, including Kniest dysplasia, Marfan syndrome, Movementdisorders, Mowat-Wilson syndrome, cystic fibrosis, Muenke syndrome,Multiple neurofibromatosis, Nance-Insley syndrome, Nance-Sweeneychondrodysplasia, Niemann-Pick disease, Noack syndrome (Pfeiffersyndrome), Osler-Weber-Rendu disease, Peutz-Jeghers syndrome, Polycystickidney disease, polyostotic fibrous dysplasia (McCune-Albrightsyndrome), Peutz-Jeghers syndrome, Prader-Labhart-Willi syndrome,hemochromatosis, primary hyperuricemia syndrome (Lesch-Nyhan syndrome),primary pulmonary hypertension, primary senile degenerative dementia,prion disease, progeria (Hutchinson Gilford Progeria Syndrome),progressive chorea, chronic hereditary (Huntington) (Huntington'sdisease), progressive muscular atrophy, spinal muscular atrophy,propionic acidemia, protoporphyria, proximal myotonic dystrophy,pulmonary arterial hypertension, PXE (pseudoxanthoma elasticum), Rb(retinoblastoma), Recklinghausen disease (neurofibromatosis type I),Recurrent polyserositis, Retinal disorders, Retinoblastoma, Rettsyndrome, RFALS type 3, Ricker syndrome, Riley-Day syndrome, Roussy-Levysyndrome, severe achondroplasia with developmental delay and acanthosisnigricans (SADDAN), Li-Fraumeni syndrome, sarcoma, breast, leukemia, andadrenal gland (SBLA) syndrome, sclerosis tuberose (tuberous sclerosis),SDAT, SED congenital (spondyloepiphyseal dysplasia congenita), SEDStrudwick (spondyloepimetaphyseal dysplasia, Strudwick type), SEDc(spondyloepiphyseal dysplasia congenita) SEMD, Strudwick type(spondyloepimetaphyseal dysplasia, Strudwick type), Shprintzen syndrome,Skin pigmentation disorders, Smith-Lemli-Opitz syndrome, South-Africangenetic porphyria (variegate porphyria), infantile-onset ascendinghereditary spastic paralysis, Speech and communication disorders,sphingolipidosis, Tay-Sachs disease, spinocerebellar ataxia, Sticklersyndrome, stroke, androgen insensitivity syndrome, tetrahydrobiopterindeficiency, beta-thalassemia, Thyroid disease Tomaculous neuropathy(hereditary neuropathy with liability to pressure palsies) TreacherCollins syndrome, Triplo X syndrome (triple X syndrome), Trisomy 21(Down syndrome), Trisomy X, VHL syndrome (von Hippel-Lindau disease),Vision impairment and blindness (Alstrom syndrome), Vrolik disease,Waardenburg syndrome, Warburg Sjo Fledelius Syndrome,Weissenbacher-Zweymuller syndrome, Wolf-Hirschhorn syndrome, WolffPeriodic disease, Weissenbacher-Zweymfiller syndrome and Xerodermapigmentosum, among others.

The term “neoplasia” or “cancer” is used throughout the specification torefer to the pathological process that results in the formation andgrowth of a cancerous or malignant neoplasm, i.e., abnormal tissue thatgrows by cellular proliferation, often more rapidly than normal andcontinues to grow after the stimuli that initiated the new growth cease.Malignant neoplasms show partial or complete lack of structuralorganization and functional coordination with the normal tissue and mostinvade surrounding tissues, metastasize to several sites, and are likelyto recur after attempted removal and to cause the death of the patientunless adequately treated. As used herein, the term neoplasia is used todescribe all cancerous disease states and embraces or encompasses thepathological process associated with malignant hematogenous, ascitic andsolid tumors. Exemplary cancers which may be treated by the presentcompounds either alone or in combination with at least one additionalanti-cancer agent include squamous-cell carcinoma, basal cell carcinoma,adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas,cancer of the bladder, bowel, breast, cervix, colon, esophagus, head,kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach;leukemias; benign and malignant lymphomas, particularly Burkitt'slymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas;myeloproliferative diseases; sarcomas, including Ewing's sarcoma,hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheralneuroepithelioma, synovial sarcoma, gliomas, astrocytomas,oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas,ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors,meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowelcancer, breast cancer, prostate cancer, cervical cancer, uterine cancer,lung cancer, ovarian cancer, testicular cancer, thyroid cancer,astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, livercancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease,Wilms' tumor and teratocarcinomas.

Additional cancers which may be treated using compounds according to thepresent invention include, for example, T-lineage Acute lymphoblasticLeukemia (T-ALL), T-lineage lymphoblastic Lymphoma (T-LL), PeripheralT-cell lymphoma, Adult T-cell Leukemia, Pre-B ALL, Pre-B Lymphomas,Large B-cell Lymphoma, Burkitts Lymphoma, B-cell ALL, Philadelphiachromosome positive ALL and Philadelphia chromosome positive CML, Theterm “bioactive agent” is used to describe an agent, other than acompound according to the present invention, which is used incombination with the present compounds as an agent with biologicalactivity to assist in effecting an intended therapy, inhibition and/orprevention/prophylaxis for which the present compounds are used.Preferred bioactive agents for use herein include those agents whichhave pharmacological activity similar to that for which the presentcompounds are used or administered and include for example, anti-canceragents, antiviral agents, especially including anti-HIV agents andanti-HCV agents, antimicrobial agents, antifungal agents, etc.

The term “additional anti-cancer agent” is used to describe ananti-cancer agent, which may be combined with compounds according to thepresent invention to treat cancer. These agents include, for example,everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib,GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107,TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457,MLN8054, PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFRinhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK-1modulator, a Bcl-2 inhibitor, an HDAC inhbitor, a c-MET inhibitor, aPARP inhibitor, a Cdk inhibitor, an EGFR TK inhibitor, an IGFR-TKinhibitor, an anti-HGF antibody, a PI3 kinase inhibitors, an AKTinhibitor, a JAK/STAT inhibitor, a checkpoint-1 or 2 inhibitor, a focaladhesion kinase inhibitor, a Map kinase kinase (mek) inhibitor, a VEGFtrap antibody, pemetrexed, erlotinib, dasatanib, nilotinib, decatanib,panitumumab, amrubicin, oregovomab, Lep-etu, nolatrexed, azd2171,batabulin, ofatumumab, zanolimumab, edotecarin, tetrandrine, rubitecan,tesmilifene, oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111,131-I-TM-601, ALT-110, BIO 140, CC 8490, cilengitide, gimatecan,IL13-PE38QQR, INO 1001, IPdR₁ KRX-0402, lucanthone, LY317615, neuradiab,vitespan, Rta 744, Sdx 102, talampanel, atrasentan, Xr 311, romidepsin,ADS-100380, sunitinib, 5-fluorouracil, vorinostat, etoposide,gemcitabine, doxorubicin, liposomal doxorubicin,5′-deoxy-5-fluorouridine, vincristine, temozolomide, ZK-304709,seliciclib; PD0325901, AZD-6244, capecitabine, L-Glutamic acid,N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-,disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan,tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, DES(diethyl stilbestrol), estradiol, estrogen, conjugated estrogen,bevacizumab, IMC-1C11, CHIR-258);3-[5-(methylsulfonylpiperadinemethyl)-indolylj-quinolone, vatalanib,AG-013736, AVE-0005, the acetate salt of [D-Ser(Bu t) 6, Azgly 10](pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu t)-Leu-Arg-Pro-Azgly-NH₂ acetate[C₅₉H₈₄N₁₈Oi₄-(C₂H₄O₂)_(X) where x=1 to 2.4], goserelin acetate,leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate,hydroxyprogesterone caproate, megestrol acetate, raloxifene,bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714;TAK-165, HKI-272, erlotinib, lapatanib, canertinib, ABX-EGF antibody,erbitux, EKB-569, PKI-166, GW-572016, lonafarnib, BMS-214662,tipifarnib; amifostine, NVP-LAQ824, suberoyl analide hydroxamic acid,valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951,aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, BacillusCalmette-Guerin (BCG) vaccine, adriamycin, bleomycin, buserelin,busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine,clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin,daunorubicin, diethylstilbestrol, epirubicin, fludarabine,fludrocortisone, fluoxymesterone, flutamide, gleevec, gemcitabine,hydroxyurea, idarubicin, ifosfamide, imatinib, leuprolide, levamisole,lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna,methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide,oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer,procarbazine, raltitrexed, rituximab, streptozocin, teniposide,testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine,13-cis-retinoic acid, phenylalanine mustard, uracil mustard,estramustine, altretamine, floxuridine, 5-deooxyuridine, cytosinearabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin,mithramycin, vinblastine, vinorelbine, topotecan, razoxin, marimastat,COL-3, neovastat, BMS-275291, squalamine, endostatin, SU5416, SU6668,EMD121974, interleukin-12, IM862, angiostatin, vitaxin, droloxifene,idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab,denileukin diftitox, gefitinib, bortezimib, paclitaxel, cremophor-freepaclitaxel, docetaxel, epithilone B, BMS-247550, BMS-310705,droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA-923, arzoxifene,fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR-3339,ZK186619, topotecan, PTK787/ZK 222584, VX-745, PD 184352, rapamycin,40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001,ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646,wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin,erythropoietin, granulocyte colony-stimulating factor, zolendronate,prednisone, cetuximab, granulocyte macrophage colony-stimulating factor,histrelin, pegylated interferon alfa-2a, interferon alfa-2a, pegylatedinterferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L-asparaginase,lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane,alemtuzumab, all-transretinoic acid, ketoconazole, interleukin-2,megestrol, immune globulin, nitrogen mustard, methylprednisolone,ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine,bexarotene, tositumomab, arsenic trioxide, cortisone, editronate,mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase,strontium 89, casopitant, netupitant, an NK-1 receptor antagonist,palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide,lorazepam, alprazolam, haloperidol, droperidol, dronabinol,dexamethasone, methylprednisolone, prochlorperazine, granisetron,ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin,epoetin alfa, darbepoetin alfa and mixtures thereof.

The term “anti-HIV agent” or “additional anti-HIV agent” includes, forexample, nucleoside reverse transcriptase inhibitors (NRTI), othernon-nucloeoside reverse transcriptase inhibitors (i.e., those which arenot representative of the present invention), protease inhibitors,fusion inhibitors, among others, exemplary compounds of which mayinclude, for example, 3TC (Lamivudine), AZT (Zidovudine), (−)-FTC, ddI(Didanosine), ddC (zalcitabine), abacavir (ABC), tenofovir (PMPA),D-D4FC (Reverset), D4T (Stavudine), Racivir, L-FddC, L-FD4C, NVP(Nevirapine), DLV (Delavirdine), EFV (Efavirenz), SQVM (Saquinavirmesylate), RTV (Ritonavir), IDV (Indinavir), SQV (Saquinavir), NFV(Nelfinavir), APV (Amprenavir), LPV (Lopinavir), fusion inhibitors suchas T20, among others, fuseon and mixtures thereof, including anti-HIVcompounds presently in clinical trials or in development.

Other anti-HIV agents which may be used in coadministration withcompounds according to the present invention include, for example, otherNNRTI's (i.e., other than the NNRTI's according to the presentinvention) may be selected from the group consisting of nevirapine(BI-R6-587), delavirdine (U-90152S/T), efavirenz (DMP-266), UC-781(N-[4-chloro-3-(3-methyl-2-butenyloxy)phenyl]-2methyl3-furancarbothiamide),etravirine (TMC125), Trovirdine (Ly300046.HCl), MKC-442 (emivirine,coactinon), HI-236, HI-240, HI-280, HI-281, rilpivirine (TMC-278),MSC-127, HBY 097, DMP266, Baicalin (TJN-151) ADAM-II (Methyl3′,3′-dichloro-4′,4″-dimethoxy-5′,5″-bis(methoxycarbonyl)-6,6-diphenylhexenoate),Methyl3-Bromo-5-(1-5-bromo-4-methoxy-3-(methoxycarbonyl)phenyl)hept-1-enyl)-2-methoxybenzoate(Alkenyldiarylmethane analog, Adam analog), 5C13PhS-2Indo1CONH2(5-chloro-3-(phenylsulfinyl)-2′-indolecarboxamide), AAP-BHAP (U-104489or PNU-104489), Capravirine (AG-1549, S-1153), atevirdine (U-87201E),aurin tricarboxylic acid (SD-095345),1-[(6-Cyano-2-indoyly)carbonyl]-4-[3-(isopropylamino)-2-pyridinyl]piperazine(piperazine1 pyridine 4 indolyl derivative),1-[5-[[N-(methyl)methylsulfonylamino]-2-indolylcarbonyl-4-[3-(isopropylamino)-2-pyridinyl]piperazine(piperazine 1pyridine 5 indolyl derivative),1-[3-(Ethylamino)-2-[pyridinyl]-4-[(5-hydroxy-2-indolyl)carbonyl]piperazine,1-[(6-Formyl-2-indoyly)carbonyl]-4-[3-(isopropylamino)-2-pyridinyl]piperazine,1-[[5-(Methylsulfonyloxy)-2-indoyly)carbonyl]-4-[3-(isopropylamino)-2-pyridinyl]piperazine,U88204E, Bis(2-nitrophenyl)sulfone (NSC 633001), Calanolide A(NSC675451), Calanolide B,6-Benzyl-5-methyl-2-(cyclohexyloxy)pyrimidin-4-one (DABO-546), DPC 961,E-EBU, E-EBU-dm, E-EPSeU, E-EPU, Foscarnet (Foscavir), HEPT(1-[(2-Hydroxyethoxy)methyl]-6-(phenylthio)thymine), HEPT-M(1-[(2-Hydroxyethoxy)methyl]-6-(3-methylphenyl)thio)thymine), HEPT-S(1-[(2-Hydroxyethoxy)methyl]-6-(phenylthio)-2-thiothymine), InophyllumP, L-737,126, Michellamine A (NSC650898), Michellamine B (NSC649324),Michellamine F,6-(3,5-Dimethylbenzyl)-1-[(2-hydroxyethoxy)methyl]-5-isopropyluracil,6-(3,5-Dimethylbenzyl)-1-(ethyoxymethyl)-5-isopropyluracil, NPPS, E-BPTU(NSC 648400), Oltipraz(4-Methyl-5-(pyrazinyl)-3H-1,2-dithiole-3-thione),N-{2-(2-Chloro-6-fluorophenethyl]-N′-(2-thiazolyl)thiourea (PETT Cl, Fderivative),N-{2-(2,6-Difluorophenethyl]-N′-[2-(5-bromopyridyl)]thiourea {PETTderivative),N-{2-(2,6-Difluorophenethyl]-N′-[2-(5-methylpyridyl)]hiourea {PETTPyridyl derivative),N-[2-(3-Fluorofuranyl)ethyl]-N′-[2-(5-chloropyridyl)]thiourea,N-[2-(2-Fluoro-6-ethoxyphenethyl)]-N′-[2-(5-bromopyridyl)]thiourea,N-(2-Phenethyl)-N′-(2-thiazolyl)thiourea (LY-73497), L-697,639,L-697,593, L-697,661,3-[2-(4,7-Difluorobenzoxazol-2-yl)ethyl}-5-ethyl-6-methyl(pypridin-2(1H)-thione(2-Pyridinone Derivative),3-[[(2-Methoxy-5,6-dimethyl-3-pyridyl)methyl]amine]-5-ethyl-6-methyl(pypridin-2(1H)-thione(2-Pyridinone 3pyrid 3MeNH Derivative), R82150, R82913, R87232, R88703,R89439 (Loviride), R90385, S-2720, Suramin Sodium, TBZ(Thiazolobenzimidazole, NSC 625487), Thiazoloisoindol-5-one,(+)(R)-9b-(3,5-Dimethylphenyl-2,3-dihydrothiazolo[2,3-a]isoindol-5(9bH)-one, Tivirapine (R86183), UC-38 and UC-84, among others.

The term “pharmaceutically acceptable salt” is used throughout thespecification to describe, where applicable, a salt form of one or moreof the compounds described herein which are presented to increase thesolubility of the compound in the gastic juices of the patient'sgastrointestinal tract in order to promote dissolution and thebioavailability of the compounds. Pharmaceutically acceptable saltsinclude those derived from pharmaceutically acceptable inorganic ororganic bases and acids, where applicable. Suitable salts include thosederived from alkali metals such as potassium and sodium, alkaline earthmetals such as calcium, magnesium and ammonium salts, among numerousother acids and bases well known in the pharmaceutical art. Sodium andpotassium salts are particularly preferred as neutralization salts ofthe phosphates according to the present invention.

The term “pharmaceutically acceptable derivative” is used throughout thespecification to describe any pharmaceutically acceptable prodrug form(such as an ester, amide other prodrug group), which, uponadministration to a patient, provides directly or indirectly the presentcompound or an active metabolite of the present compound.

The term “independently” is used herein to indicate that the variable,which is independently applied, varies independently from application toapplication.

The term “hydrocarbyl” shall mean a compound which contains carbon andhydrogen and which may be fully saturated, partially unsaturated oraromatic and includes aryl groups, alkyl groups, alkenyl groups andalkynyl groups.

The term “alkyl” shall mean within its context a linear, branch-chainedor cyclic fully saturated hydrocarbon radical or alkyl group, preferablya C₁-C₁₀, more preferably a C₁-C₆, alternatively a C₁-C₃ alkyl group,which may be optionally substituted. Examples of alkyl groups aremethyl, ethyl, n-butyl, sec-butyl, n-hexyl, n-heptyl, n-octyl, n-nonyl,n-decyl, isopropyl, 2-methylpropyl, cyclopropyl, cyclopropylmethyl,cyclobutyl, cyclopentyl, cyclopentylethyl, cyclohexylethyl andcyclohexyl, among others. In certain preferred embodiments, compoundsaccording to the present invention which may be used to covalently bindto dehalogenase enzymes. These compounds generally contain a side chain(often linked through a polyethylene glycol group) which terminates inan alkyl group which has a halogen substituent (often chlorine orbromine) on its distil end which results in covalent binding of thecompound containing such a moiety to the protein. The term “Alkenyl”refers to linear, branch-chained or cyclic C₂-C₁₀ (preferably C₂-C₆)hydrocarbon radicals containing at least one C═C bond. The term“Alkynyl” refers to linear, branch-chained or cyclic C₂-C₁₀ (preferablyC₂-C₆) hydrocarbon radicals containing at least one C≡C bond. The term“alkylene” when used, refers to a —(CH₂)_(n)— group (n is an integergenerally from 0-6), which may be optionally substituted. Whensubstituted, the alkylene group preferably is substituted on one or moreof the methylene groups with a C₁-C₆ alkyl group (including acyclopropyl group or a t-butyl group), more preferably a methyl group,but may also be substituted with one or more halo groups, preferablyfrom 1 to 3 halo groups or one or two hydroxyl groups, O—(C₁-C₆ alkyl)groups or amino acid sidechains as otherwise disclosed herein. Incertain embodiments, an alkylene group may be substituted with aurethane or alkoxy group (or other group) which is further substitutedwith a polyethylene glycol chain (of from 1 to 10, preferably 1 to 6,often 1 to 4 ethylene glycol units) to which is substituted (preferably,but not exclusively on the distal end of the polyethylene glycol chain)an alkyl chain substituted with a single halogen group, preferably achlorine group. In still other embodiments, the alkylene (often, amethylene) group, may be substituted with an amino acid sidechain groupsuch as a sidechain group of a natural or unnatural amino acid, forexample, alanine, β-alanine, arginine, asparagine, aspartic acid,cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine,histidine, isoleucine, lysine, leucine, methionine, proline, serine,threonine, valine, tryptophan or tyrosine.

The term “unsubstituted” shall mean substituted only with hydrogenatoms. A range of carbon atoms which includes C₀ means that carbon isabsent and is replaced with H. Thus, a range of carbon atoms which isC₀-C₆ includes carbons atoms of 1, 2, 3, 4, 5 and 6 and for C₀, H standsin place of carbon. The term “substituted” or “optionally substituted”shall mean independently (i.e., where more than substituent occurs, eachsubstituent is independent of another substituent) one or moresubstituents (independently up to five substitutents, preferably up tothree substituents, often 1 or 2 substituents on a moiety in a compoundaccording to the present invention and may include substituents whichthemselves may be further substituted) at a carbon (or nitrogen)position anywhere on a molecule within context, and includes assubstituents hydroxyl, thiol, carboxyl, cyano (C≡N), nitro (NO₂),halogen (preferably, 1, 2 or 3 halogens, especially on an alkyl,especially a methyl group such as a trifluoromethyl), an alkyl group(preferably, C₁-C₁₀, more preferably, C₁-C₆), aryl (especially phenyland substituted phenyl for example benzyl or benzoyl), alkoxy group(preferably, C₁-C₆ alkyl or aryl, including phenyl and substitutedphenyl), thioether (C₁-C₆ alkyl or aryl), acyl (preferably, C₁-C₆ acyl),ester or thioester (preferably, C₁-C₆ alkyl or aryl) including alkyleneester (such that attachment is on the alkylene group, rather than at theester function which is preferably substituted with a C₁-C₆ alkyl oraryl group), preferably, C₁-C₆ alkyl or aryl, halogen (preferably, F orCl), amine (including a five- or six-membered cyclic alkylene amine,further including a C₁-C₆ alkyl amine or a C₁-C₆ dialkyl amine whichalkyl groups may be substituted with one or two hydroxyl groups) or anoptionally substituted —N(C₀-C₆ alkyl)C(O)(O—C₁-C₆ alkyl) group (whichmay be optionally substituted with a polyethylene glycol chain to whichis further bound an alkyl group containing a single halogen, preferablychlorine substituent), hydrazine, amido, which is preferably substitutedwith one or two C₁-C₆ alkyl groups (including a carboxamide which isoptionally substituted with one or two C₁-C₆ alkyl groups), alkanol(preferably, C₁-C₆ alkyl or aryl), or alkanoic acid (preferably, C₁-C₆alkyl or aryl). Substituents according to the present invention mayinclude, for example —SiR₁R₂R₃ groups where each of R₁ and R₂ is asotherwise described herein and R₃ is H or a C₁-C₆ alkyl group,preferably R₁, R₂, R₃ in this context is a C₁-C₃ alkyl group (includingan isopropyl or t-butyl group). Each of the above-described groups maybe linked directly to the substituted moiety or alternatively, thesubstituent may be linked to the substituted moiety (preferably in thecase of an aryl or heteraryl moiety) through an optionally substituted—(CH₂)_(m)— or alternatively an optionally substituted —(OCH₂)_(m)—,—(OCH₂CH₂)_(m)— or —(CH₂CH₂O)_(m)— group, which may be substituted withany one or more of the above-described substituents. Alkylene groups—(CH₂)_(m)— or —(CH₂)_(n)— groups or other chains such as ethyleneglycol chains, as identified above, may be substituted anywhere on thechain. Preferred substitutents on alkylene groups include halogen orC₁-C₆ (preferably C₁-C₃) alkyl groups, which may be optionallysubstituted with one or two hydroxyl groups, one or two ether groups(0-C₁-C₆ groups), up to three halo groups (preferably F), or a sideshainof an amino acid as otherwise described herein and optionallysubstituted amide (preferably carboxamide substituted as describedabove) or urethane groups (often with one or two C₀-C₆ alkylsubstitutents, which group(s) may be further substituted). In certainembodiments, the alkylene group (often a single methylene group) issubstituted with one or two optionally substituted C₁-C₆ alkyl groups,preferably C₁-C₄ alkyl group, most often methyl or O-methyl groups or asidechain of an amino acid as otherwise described herein. In the presentinvention, a moiety in a molecule may be optionally substituted with upto five substituents, preferably up to three substituents. Most often,in the present invention moieties which are substituted are substitutedwith one or two substituents.

The term “substituted” (each substituent being independent of any othersubstituent) shall also mean within its context of use C₁-C₆ alkyl,C₁-C₆ alkoxy, halogen, amido, carboxamido, sulfone, includingsulfonamide, keto, carboxy, C₁-C₆ ester (oxyester or carbonylester),C₁-C₆ keto, urethane —O—C(O)—NR₁R₂ or —N(R₁)—C(O)—O—R₁, nitro, cyano andamine (especially including a C₁-C₆ alkylene-NR₁R₂, a mono- or di-C₁-C₆alkyl substituted amines which may be optionally substituted with one ortwo hydroxyl groups). Each of these groups contain unless otherwiseindicated, within context, between 1 and 6 carbon atoms. In certainembodiments, preferred substituents will include for example, —NH—,—NHC(O)—, —O—, ═O, —(CH₂)_(m)— (here, m and n are in context, 1, 2, 3,4, 5 or 6), —S—, —S(O)—, SO₂— or —NH—C(O)—NH—, —(CH₂)_(n)OH,—(CH₂)_(n)SH, —(CH₂)_(n)COOH, C₁-C₆ alkyl, —(CH₂)_(n)O—(C₁-C₆ alkyl),—(CH₂)_(n)C(O)—(C₁-C₆ alkyl), —(CH₂)_(n)OC(O)—(C₁-C₆ alkyl),—(CH₂)_(n)C(O)—(C₁-C₆ alkyl), —(CH₂)_(n)NHC(O)—R₁, —(CH₂)_(n)C(O)—NR¹R₂,—(OCH₂)_(n)OH, —(CH₂O)_(n)COOH, C₁-C₆ alkyl, —(OCH₂)_(n)O—(C₁-C₆ alkyl),—(CH₂O)_(n)C(O)—(C₁-C₆ alkyl), —(OCH₂)_(n)NHC(O)—R₁,—(CH₂O)_(n)C(O)—NR₁R₂, —S(O)₂—R_(S), —S(O)—R_(S) (R_(S) is C₁-C₆ alkylor a —(CH₂)_(m)—NR₁R₂ group), NO₂, CN or halogen (F, Cl, Br, I,preferably F or Cl), depending on the context of the use of thesubstituent. R₁ and R₂ are each, within context, H or a C₁-C₆ alkylgroup (which may be optionally substituted with one or two hydroxylgroups or up to three halogen groups, preferably fluorine). The term“substituted” shall also mean, within the chemical context of thecompound defined and substituent used, an optionally substituted aryl orheteroaryl group or an optionally substituted heterocyclic group asotherwise described herein. Alkylene groups may also be substituted asotherwise disclosed herein, preferably with optionally substituted C₁-C₆alkyl groups (methyl, ethyl or hydroxymethyl or hydroxyethyl ispreferred, thus providing a chiral center), a sidechain of an amino acidgroup as otherwise described herein, an amido group as describedhereinabove, or a urethane group O—C(O)—NR₁R₂ group where R₁ and R₂ areas otherwise described herein, although numerous other groups may alsobe used as substituents. Various optionally substituted moieties may besubstituted with 3 or more substituents, preferably no more than 3substituents and preferably with 1 or 2 substituents. It is noted thatin instances where, in a compound at a particular position of themolecule substitution is required (principally, because of valency), butno substitution is indicated, then that substituent is construed orunderstood to be H, unless the context of the substitution suggestsotherwise.

The term “aryl” or “aromatic”, in context, refers to a substituted (asotherwise described herein) or unsubstituted monovalent aromatic radicalhaving a single ring (e.g., benzene, phenyl, benzyl) or condensed rings(e.g., naphthyl, anthracenyl, phenanthrenyl, etc.) and can be bound tothe compound according to the present invention at any available stableposition on the ring(s) or as otherwise indicated in the chemicalstructure presented. Other examples of aryl groups, in context, mayinclude heterocyclic aromatic ring systems “heteroaryl” groups havingone or more nitrogen, oxygen, or sulfur atoms in the ring (moncyclic)such as imidazole, furyl, pyrrole, furanyl, thiene, thiazole, pyridine,pyrimidine, pyrazine, triazole, oxazole or fused ring systems such asindole, quinoline, indolizine, azaindolizine, benzofurazan, etc., amongothers, which may be optionally substituted as described above. Amongthe heteroaryl groups which may be mentioned include nitrogen-containingheteroaryl groups such as pyrrole, pyridine, pyridone, pyridazine,pyrimidine, pyrazine, pyrazole, imidazole, triazole, triazine,tetrazole, indole, isoindole, indolizine, azaindolizine, purine,indazole, quinoline, dihydroquinoline, tetrahydroquinoline,isoquinoline, dihydroisoquinoline, tetrahydroisoquinoline, quinolizine,phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline,pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine,acridine, phenanthridine, carbazole, carbazoline, perimidine,phenanthroline, phenacene, oxadiazole, benzimidazole, pyrrolopyridine,pyrrolopyrimidine and pyridopyrimidine; sulfur-containing aromaticheterocycles such as thiophene and benzothiophene; oxygen-containingaromatic heterocycles such as furan, pyran, cyclopentapyran, benzofuranand isobenzofuran; and aromatic heterocycles comprising 2 or more heteroatoms selected from among nitrogen, sulfur and oxygen, such as thiazole,thiadizole, isothiazole, benzoxazole, benzothiazole, benzothiadiazole,phenothiazine, isoxazole, furazan, phenoxazine, pyrazoloxazole,imidazothiazole, thienofuran, furopyrrole, pyridoxazine, furopyridine,furopyrimidine, thienopyrimidine and oxazole, among others, all of whichmay be optionally substituted.

The term “heterocycle” refers to a cyclic group which contains at leastone heteroatom, i.e., O, N or S, and may be aromatic (heteroaryl) ornon-aromatic. Thus, the heteroaryl moieties are subsumed under thedefinition of heterocycle, depending on the context of its use.Exemplary heteroaryl groups are described hereinabove. Exemplarynon-aromatic heterocyclic groups for use in the present inventioninclude, for example, pyrrolidinyl, pyrrolinyl, piperidinyl,piperazinyl, N-methylpiperazinyl, pyrazolidinyl, imidazolidinyl,morpholinyl, tetrahydropyranyl, azetidinyl, oxetanyl, oxathiolanyl,pyridone, 2-pyrrolidone, ethyleneurea, 1,3-dioxolane, 1,3-dioxane,1,4-dioxane, phthalimide and succinimide, among others, as describedherein.

The terms “treat”, “treating”, and “treatment”, etc., as used herein,refer to any action providing a benefit to a patient for which thepresent compounds may be administered, including the treatment of anydisease state or condition which is modulated through the protein towhich the present compounds bind. Disease states or conditions,including cancer, which may be treated using compounds according to thepresent invention are set forth hereinabove.

The term “coadministration” or “combination therapy” shall mean that atleast two compounds or compositions are administered to the patient atthe same time, such that effective amounts or concentrations of each ofthe two or more compounds may be found in the patient at a given pointin time. Although compounds according to the present invention may beco-administered to a patient at the same time, the term embraces bothadministration of two or more agents at the same time or at differenttimes, provided that effective concentrations of all coadministeredcompounds or compositions are found in the subject at a given time. Incertain preferred aspects of the present invention, one or more of thepresent compounds described above, are coadministered in combinationwith at least one additional bioactive agent, especially including ananticancer agent. In particularly preferred aspects of the invention,the co-administration of compounds results in synergistic therapeutic,including anticancer therapy.

Pharmaceutical compositions comprising combinations of an effectiveamount of at least one bifunctional compound according to the presentinvention, and one or more of the compounds otherwise described herein,all in effective amounts, in combination with a pharmaceuticallyeffective amount of a carrier, additive or excipient, represents afurther aspect of the present invention.

The present invention includes, where applicable, the compositionscomprising the pharmaceutically acceptable salts, in particular, acid orbase addition salts of compounds of the present invention. The acidswhich are used to prepare the pharmaceutically acceptable acid additionsalts of the aforementioned base compounds useful in this invention arethose which form non-toxic acid addition salts, i.e., salts containingpharmacologically acceptable anions, such as the hydrochloride,hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acidphosphate, acetate, lactate, citrate, acid citrate, tartrate,bitartrate, succinate, maleate, fumarate, gluconate, saccharate,benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate,p-toluenesulfonate and pamoate [i.e., 1,1′-methylene-bis-(2-hydroxy-3naphthoate)]salts, among numerous others.

Pharmaceutically acceptable base addition salts may also be used toproduce pharmaceutically acceptable salt forms of the compounds orderivatives according to the present invention. The chemical bases thatmay be used as reagents to prepare pharmaceutically acceptable basesalts of the present compounds that are acidic in nature are those thatform non-toxic base salts with such compounds. Such non-toxic base saltsinclude, but are not limited to those derived from suchpharmacologically acceptable cations such as alkali metal cations (eg.,potassium and sodium) and alkaline earth metal cations (eg, calcium,zinc and magnesium), ammonium or water-soluble amine addition salts suchas N-methylglucamine-(meglumine), and the lower alkanolammonium andother base salts of pharmaceutically acceptable organic amines, amongothers.

The compounds of the present invention may, in accordance with theinvention, be administered in single or divided doses by the oral,parenteral or topical routes. Administration of the active compound mayrange from continuous (intravenous drip) to several oral administrationsper day (for example, Q.I.D.) and may include oral, topical, parenteral,intramuscular, intravenous, sub-cutaneous, transdermal (which mayinclude a penetration enhancement agent), buccal, sublingual andsuppository administration, among other routes of administration.Enteric coated oral tablets may also be used to enhance bioavailabilityof the compounds from an oral route of administration. The mosteffective dosage form will depend upon the pharmacokinetics of theparticular agent chosen as well as the severity of disease in thepatient. Administration of compounds according to the present inventionas sprays, mists, or aerosols for intra-nasal, intra-tracheal orpulmonary administration may also be used. The present inventiontherefore also is directed to pharmaceutical compositions comprising aneffective amount of compound according to the present invention,optionally in combination with a pharmaceutically acceptable carrier,additive or excipient. Compounds according to the present invention maybe administered in immediate release, intermediate release or sustainedor controlled release forms. Sustained or controlled release forms arepreferably administered orally, but also in suppository and transdermalor other topical forms. Intramuscular injections in liposomal form mayalso be used to control or sustain the release of compound at aninjection site.

The compositions of the present invention may be formulated in aconventional manner using one or more pharmaceutically acceptablecarriers and may also be administered in controlled-releaseformulations. Pharmaceutically acceptable carriers that may be used inthese pharmaceutical compositions include, but are not limited to, ionexchangers, alumina, aluminum stearate, lecithin, serum proteins, suchas human serum albumin, buffer substances such as phosphates, glycine,sorbic acid, potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as prolaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

The compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously.

Sterile injectable forms of the compositions of this invention may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1, 3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of inj ectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such as Ph. Helv orsimilar alcohol.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers which are commonly used includelactose and corn starch. Lubricating agents, such as magnesium stearate,are also typically added. For oral administration in a capsule form,useful diluents include lactose and dried corn starch. When aqueoussuspensions are required for oral use, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions of this invention may beadministered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient, which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions of this invention may also beadministered topically. Suitable topical formulations are readilyprepared for each of these areas or organs. Topical application for thelower intestinal tract can be effected in a rectal suppositoryformulation (see above) or in a suitable enema formulation.Topically-acceptable transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. In certain preferred aspects of the invention, the compounds maybe coated onto a stent which is to be surgically implanted into apatient in order to inhibit or reduce the likelihood of occlusionoccurring in the stent in the patient.

Alternatively, the pharmaceutical compositions can be formulated in asuitable lotion or cream containing the active components suspended ordissolved in one or more pharmaceutically acceptable carriers. Suitablecarriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith our without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

The amount of compound in a pharmaceutical composition of the instantinvention that may be combined with the carrier materials to produce asingle dosage form will vary depending upon the host and diseasetreated, the particular mode of administration. Preferably, thecompositions should be formulated to contain between about 0.05milligram to about 750 milligrams or more, more preferably about 1milligram to about 600 milligrams, and even more preferably about 10milligrams to about 500 milligrams of active ingredient, alone or incombination with at least one other compound according to the presentinvention.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease or condition beingtreated.

A patient or subject in need of therapy using compounds according to thepresent invention can be treated by administering to the patient(subject) an effective amount of the compound according to the presentinvention including pharmaceutically acceptable salts, solvates orpolymorphs, thereof optionally in a pharmaceutically acceptable carrieror diluent, either alone, or in combination with other knownerythopoiesis stimulating agents as otherwise identified herein.

These compounds can be administered by any appropriate route, forexample, orally, parenterally, intravenously, intradermally,subcutaneously, or topically, including transdermally, in liquid, cream,gel, or solid form, or by aerosol form.

The active compound is included in the pharmaceutically acceptablecarrier or diluent in an amount sufficient to deliver to a patient atherapeutically effective amount for the desired indication, withoutcausing serious toxic effects in the patient treated. A preferred doseof the active compound for all of the herein-mentioned conditions is inthe range from about 10 ng/kg to 300 mg/kg, preferably 0.1 to 100 mg/kgper day, more generally 0.5 to about 25 mg per kilogram body weight ofthe recipient/patient per day. A typical topical dosage will range from0.01-5% wt/wt in a suitable carrier.

The compound is conveniently administered in any suitable unit dosageform, including but not limited to one containing less than 1 mg, 1 mgto 3000 mg, preferably 5 to 500 mg of active ingredient per unit dosageform. An oral dosage of about 25-250 mg is often convenient.

The active ingredient is preferably administered to achieve peak plasmaconcentrations of the active compound of about 0.00001-30 mM, preferablyabout 0.1-30 μM. This may be achieved, for example, by the intravenousinjection of a solution or formulation of the active ingredient,optionally in saline, or an aqueous medium or administered as a bolus ofthe active ingredient. Oral administration is also appropriate togenerate effective plasma concentrations of active agent.

The concentration of active compound in the drug composition will dependon absorption, distribution, inactivation, and excretion rates of thedrug as well as other factors known to those of skill in the art. It isto be noted that dosage values will also vary with the severity of thecondition to be alleviated. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions, and that the concentration ranges set forth herein areexemplary only and are not intended to limit the scope or practice ofthe claimed composition. The active ingredient may be administered atonce, or may be divided into a number of smaller doses to beadministered at varying intervals of time.

Oral compositions will generally include an inert diluent or an ediblecarrier. They may be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound or its prodrug derivative can be incorporated with excipientsand used in the form of tablets, troches, or capsules. Pharmaceuticallycompatible binding agents, and/or adjuvant materials can be included aspart of the composition.

The tablets, pills, capsules, troches and the like can contain any ofthe following ingredients, or compounds of a similar nature: a bindersuch as microcrystalline cellulose, gum tragacanth or gelatin; anexcipient such as starch or lactose, a dispersing agent such as alginicacid, Primogel, or corn starch; a lubricant such as magnesium stearateor Sterotes; a glidant such as colloidal silicon dioxide; a sweeteningagent such as sucrose or saccharin; or a flavoring agent such aspeppermint, methyl salicylate, or orange flavoring. When the dosage unitform is a capsule, it can contain, in addition to material of the abovetype, a liquid carrier such as a fatty oil. In addition, dosage unitforms can contain various other materials which modify the physical formof the dosage unit, for example, coatings of sugar, shellac, or entericagents.

The active compound or pharmaceutically acceptable salt thereof can beadministered as a component of an elixir, suspension, syrup, wafer,chewing gum or the like. A syrup may contain, in addition to the activecompounds, sucrose as a sweetening agent and certain preservatives, dyesand colorings and flavors.

The active compound or pharmaceutically acceptable salts thereof canalso be mixed with other active materials that do not impair the desiredaction, or with materials that supplement the desired action, such aserythropoietin stimulating agents, including EPO and darbapoietin alfa,among others. In certain preferred aspects of the invention, one or morecompounds according to the present invention are coadministered withanother bioactive agent, such as an erythropoietin stimulating agent ora would healing agent, including an antibiotic, as otherwise describedherein.

Solutions or suspensions used for parenteral, intradermal, subcutaneous,or topical application can include the following components: a sterilediluent such as water for injection, saline solution, fixed oils,polyethylene glycols, glycerine, propylene glycol or other syntheticsolvents; antibacterial agents such as benzyl alcohol or methylparabens; antioxidants such as ascorbic acid or sodium bisulfite;chelating agents such as ethylenediaminetetraacetic acid; buffers suchas acetates, citrates or phosphates and agents for the adjustment oftonicity such as sodium chloride or dextrose. The parental preparationcan be enclosed in ampoules, disposable syringes or multiple dose vialsmade of glass or plastic.

If administered intravenously, preferred carriers are physiologicalsaline or phosphate buffered saline (PBS).

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art.

Liposomal suspensions may also be pharmaceutically acceptable carriers.These may be prepared according to methods known to those skilled in theart, for example, as described in U.S. Pat. No. 4,522,811 (which isincorporated herein by reference in its entirety). For example, liposomeformulations may be prepared by dissolving appropriate lipid(s) (such asstearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline,arachadoyl phosphatidyl choline, and cholesterol) in an inorganicsolvent that is then evaporated, leaving behind a thin film of driedlipid on the surface of the container. An aqueous solution of the activecompound are then introduced into the container. The container is thenswirled by hand to free lipid material from the sides of the containerand to disperse lipid aggregates, thereby forming the liposomalsuspension.

General Synthetic Approaches

Generic scheme for the synthesis of ULM derivatives is described here.Briefly, the compounds according to the present invention aresynthesized pursuant to the general solution phase synthetic scheme(presented hereinbelow) and/or general scheme I, which is directed tophase synthesis of compounds according to the present invention.Initially a hydroxyl-protected carboxy substituted (and protected)pyrrolidine compound is reacted with a carboxylic acid containingreagent, which introduces a carbonyl group at the amine of thepyrollidine ring to form an amide group. Alternatively, the pyrrolidineamine may be protected and the carboxylic acid moiety may be condensedwith a nucleophilic group on a right hand fragment to provide an amideon the right hand portion of the pyrrolidine moiety. The left and righthand fragments to be condensed onto, respectively, the amine andcarboxylic acid group of the pyrrolidine moiety are preferably preparedprior to condensing onto the pyrrolidine group, but other approaches maybe taken to introduce groups onto the pyrrolidine group. The individualcomponents which are combined to produce a ULM group can be preparedusing blocking groups at preferred functional groups on the ULM groupwhich can be removed so as to react with and covalently link a linkergroup which is prepared to accommodate a PTM moiety to which is alreadybound a protein binding moiety or PTM group or may be further reacted toform a covalent bound with a PTM group, which may also may comprise aULM′ group as otherwise described herein. Thus, a carboxylic acidcontaining left hand fragment may be condensed onto the amine group ofthe pyrroline, thus forming an amide group with an R¹ left hand fragmentas depicted below. Onto the carboxyl group, any number of nucleophilic(preferably, amine containing) right hand fragments (pre-synthesized)may be condensed onto the carboxyl group to provide an amide group withan R² right fragment as depicted below. Formation of the pre-synthesizedgroups to condense onto the amine and/or the carboxyl moiety of thepyrrolidine proceeds in a facile manner. Virtually any compound can besynthesized readily using this approach. The solid phase syntheticmethod can also be used and employs similar methods used in the solutionphase synthesis, the major difference being that the hydroxyl group maybe bound to a solid support as the other steps of the synthesis occur.The general synthetic methods are applicable to virtually all of thecompounds of the present invention with facile modifications being madeconsistent with the state of chemical synthetic art as used directly oradapted from the specific teachings of the examples which follow.

Alternative General Method for Solid Phase Synthesis of VHL Ligandsaccording to the present invention (details for which are presented inthe second set of examples), set forth herein:

Synthetic Approaches for Compound Generation to Screen for TargetProtein Binding Elements (PTM) and Ubiquitination Ligand Moieties (ULM)of the Subject Invention

Two basic methods which are used in combinatorial chemistry to identifyPTM moieties and ULM moieties are solid-phase and solution-phasemethods. Using these methods combinatorial compounds are created eitherby solution-phase synthesis or by producing compounds bound covalentlyto solid-phase particles. Once their moieties are identified they may bemodified using appropriate groups (electrophilic and/or nucleophilic),and condensed onto linker groups to produce bifunctional compoundsaccording to the present invention.

Solid Phase Methods

Solid Phase Methods rely on the teachings of Fruchtel, et al. 1996,Angew. Chem. Int. Ed. Engl. 35, 17-42; which is incorporated byreference in its entirety herein).

Solid-phase synthesis makes it easier to conduct multistep reactions andto drive reactions to completion, because excess reagents can be addedand then easily washed away after each reaction step. Another key factorin favor of solid-phase synthesis is that it makes it possible to usesplit synthesis, a technique developed in 1982. Split synthesis produceslarge support-bound libraries in which each solid-phase particle holds asingle compound, or soluble libraries produced by cleavage of compoundsfrom the solid support. For example in a split synthesis method if youhave 3 compound addition steps with 10 compounds used at each step i.e.10 containers for those compounds. This will generate 10³ compounds.Also, if you consider all the reaction steps included in a synthesis10,000 compounds made via a solid phase methods using a three-stepchemistry may only require about 22 containers for the chemistry andabout 66 liquid handling steps relative to the 10,000 containers and30,000 liquid handling steps. When you combine these advantages of solidphase synthesis with split synthesis, a significant level of synergy isachieved.

Solution Phase Methods

Solution phase chemistry is favored by many for library construction dueto the wider range of organic reactions available for solution-phasesynthesis, the technology used traditionally by most synthetic organicchemists, and products in solution can be more easily identified instandard drug target assays and characterized. A problem forsolution-phase synthesis of one molecule at a time is the finalpurification that can be both expensive and slow. Chromatography iscommonly a first resort since it usually works. In addition, theproblems associated with solution chemistry are compounded whenattempting to make tens of thousands of compounds to generate a libraryor a ‘book’ for a library.

In the generation of libraries of compounds numerous methods have beendevised resulting in the wide spread use of large libraries of chemicalsto readily allow the discovery of potential drug candidates. Thegeneration of chemical libraries that are free in solution is typicallythe goal of most of the pharmaceutical industry. This aim is due to thenature of many of the drug targets and the associated assays. Also theconstruction and utility of chemical libraries is typically facilitatedbut the generation of master plates of compounds in solution to form thebasis of the chemical library. Thus the general advantages of the solidphase synthesis methods are typically not fully realized in the contextof the current drug discovery efforts. The main reason for this is theinterest not in binding of the compound to the drug target but todemonstrate that the activity of the drug target is altered, whichtypically requires compound free in solution. Further concerns withlibraries of compounds on a solid phase arise from concerns of thepotential influence of the linker and steric effects on the compoundsbound to the solid phase.

Thus methods for the discovery of compounds, which bind to targetmolecules is known in the art. Also, the optimization of the initiallydiscovered compound is well known in the art where the affinity isimproved by generation of a pool of related compound via a moreselective combinatorial chemistry approach.

The present invention provides a mechanism to overcome these problems indrug and small molecule discovery.

Addition of the Ubiquitin Ligase Binding Moiety (ULM)

At this point in the compound discovery path for the subject invention,the target protein-binding element of the compounds of the invention hasbeen identified. These optimal binding molecules are then subjected tofurther chemistry to add the ubiquitin ligase binding moiety (ULM),pursuant to the disclosure of the present application.

An alternative approach to the discovery of the target protein-bindingmoiety is based on solution phase screening. In such an examplecompounds (available either via synthesis, natural products or fromcompanies such as ArQule (www.arqule.com), Pharmacopeia(www.pharmacopiea), and Cerep (www.cerep.com) are obtained and added tothe target protein of interest and then subjected to size exclusion toremove the unbound compounds. The protein bound fraction is thensubjected to GC/MS to identify the molecules. In this way the solutionphase screening is made rapid and facile for compounds in solution.There are numerous additional ways to determine ligand binding,including, for example, detecting changes in the Tm of the protein uponthe ligand binding, among others.

Screening for Target Protein Binding Elements

Initially a target protein is selected, for example, an enzyme orprotein involved in a particular biological process. Target protein forthe subject invention come from numerous fields where small moleculesare used to achieve modulation of a biological system in eukaryoticorganisms. Examples of such fields are antivirals, antimicrobials,anti-parasitics, or other drug targets in a human patient, which may berather diverse, etc.

The target protein is then either purified from a natural source inorder to provide sufficient material for the screen or expressed viarecombinant methods to provide sufficient material for the screens.

The target protein is then either labeled directly with a detectablespecies such as a radioactive, electrochemiluminescent, andchemiluminescent or fluorescent label or with an indirectly detectablespecies such as an enzyme, or particle. Alternatively an antibody orequivalent with binding activity to the target protein is labeled.

The next step is to provide a library of compounds for screening. Alibrary of from 1,000 to 1,000,000 is typical of the size that isscreened. These are available from a series of companies, which are wellknown in the art. These libraries of compounds are used to screen forthe binding of the target protein. Ideally compounds are bought stillbound to the solid phase or are screened for binding directly toimmobilized target protein using methods as described below forscreening.

It is also possible to generate a chemical library of various potentialbinding molecules bound to a solid phase following conventional methodsto give rise to differing potential compounds. The optimal methods forthe construction of the chemical library is to employ the methods ofsplit synthesis coupled to the solid phase (as outlined above). Thelibrary is generated using a series of solid phase chemistries such asto give rise to various compilations that form the basis of a library.The library is screened in the form of a library or in the form of thecompilations. Typically one would take the products from the splitsynthesis and pool the solid phase and use this as the basis for thescreen.

To the pool of beads used as the solid phase for the synthesis, amixture of buffer, detergents, salts and blocking agents such as serumalbumin or other proteins are added. This buffer addition step is usedto block the beads or solid phase in such a way that any significantnon-specific binding of the selected target protein does not occur.Following this blocking step the beads are washed and followed by theaddition of the target protein either labeled or not. The beads or solidphase are then incubated to allow the binding of the target proteinbinding elements to the target protein. Following the incubation of thetarget molecule to the beads or solid phase the beads are washed andthen the binding of the labeled target protein detected directly. In analternative format, if the target protein is labeled with an indirectlydetectable label such as an enzyme, the beads are then placed in to asubstrate reaction solution to detect the presence of the enzyme label.In the case of an enzyme label, substrates for these detection methodsare based on insoluble chromogenic products. In the case where thetarget protein is not labeled and an antibody or equivalent isavailable, the beads are subjected to another binding reaction where theantibody or equivalent, is labeled either directly or indirectly assuggested for the labeling of the target protein. It is also possible atthis step to not use a labeled antibody or equivalent and to add afurther step where the labeled antibody or equivalent is used. Theseadditional steps can be detected using the same standard methods knownin the art as suggested for the directly labeled target protein.

Following these steps a series of beads are identified and these beadsare selected from the bead population and subject to analysis todetermine the structure of the binding molecule that is able to bind thetarget protein as in this example. This is achieved by the use of GC/MSor via molecular tags used during the construction of the library asdescribed earlier. Alternatively, a pool which was positive is re-madegenerating a series of sub pools for screening and further re-synthesisand dividing out of the various pooled compounds until a single compoundis presented in a single well for analysis allowing the determination ofthe active compound.

This method can be repeated and/or adapted for identifying peptidetarget binding moieties (PTM) for virtually any target protein.

Screening for Binding Molecules from Chemical Libraries

The step of screening for specific molecules is made easy in theinvention as only binding activity is desired and not specificmodulation of the target protein as is required in traditional drugdiscovery.

One can buy a library of compounds for screening. A library of from1,000 to 1,000,000 is typical of the size that might be screened. Theseare available from a number of companies. These libraries of compoundsare used to screen for the binding of the target protein. Ideally,compounds are purchased still bound to the solid phase or are screenedfor binding directly to immobilized target protein using methods asdescribed below for screening.

It is also possible to generate a library of from 1,000 to 100,000compounds contained on a solid phase using split synthesis methods asdescribed earlier. The library may be constructed using a series ofchemical methods resulting in pools of the solid phase used duringsynthesis, which form the basis of the entries which make up thelibrary. In addition at the final chemical coupling step used toconstruct the various entries the solid phase pools are stored insub-pools in the libraries. These so called entries and sub-pools formthe basis for screening as they contain not only pools of compounds butalso a known chemical-coupling step used in synthesis.

The library can then be screened using two approaches. In both cases thesolid phase from the chemical library to be screened is subjectedincubation with assay buffers with blocking agents such as for example;proteins (i.e. BSA, gelatin), polyvinylpyrrolidone, ficoll, heparin,detergents (i.e. SDS, Tween, NP40, Triton X-100). This incubation stepis to block the non-specific binding sites on the solid phase used inthe generation of the library and allow the determination of specificbinding events. This initial incubation is an art recognized step invarious binding assays such as ELISA, southerns, westerns etc. Followingthis incubation with blocking agents the protein of interest is thenadded to a buffer which typically has the same composition as thatduring the blocking step but can also be modified using lower or noadditional blocking agents with the exception of the detergents whichare typically always present during a binding reaction.

In one of the screening methods the entries following the blocking stepare then subjected to binding with the purified target protein. Thesolid phase from this incubation is then washed and subjected to asecond binding step with a labeled reagent which binds to the tagsequence added to the receptor sub-unit during the recombinantengineering for the expression of the receptor sub-unit. Typically anantibody to this tag recognizes the tag sequence; examples that are incommon use are the myc, flag, and his epitopes. Following the incubationwith the tag specific binding species the presence of the labeledbinding species is detected by the presence of the label that istypically an enzyme such as alkaline phosphatase or peroxidase. Thedetection step typically makes use of an insoluble chromogenic substratethat is readily detected by eye or by image analysis systems.

In an alternative method soluble substrates can also be used andscreened using ELISA plate readers, eye or other spectrophotometricmethods. In its simplest form the various entries from the library arescreened by eye to look for beads that have developed a color due to theenzymatic action on the chromogenic substrate. These colored beadsindicate that the receptor subunit is binding to one of the compoundswithin the group of entries the next step is to determine if these socalled positive group of entries contain specific binding or if bindingis just to the tag binding reagent or some non-specific activation ofthe chromogenic substrate. To achieve this, the positive entries arescreened with out the specific binding step to the receptor sub-unit. Ifthese positive entries now become negative or show significantly reducedsignals interms of positive solid phases with in the mixture then theseare considered to be real positive hits in the screen. These realpositive entries are then subjected to re-synthesis. In thisre-synthesis the initial chemical steps to create the specific bindingmolecule is unknown only the last chemical coupling step in the compoundsynthesis is know, as this formed the last chemical step whichconstructed the group of entries. During the re-synthesis of thepositive chapter the chemical step prior to the last chemical couplingis carried out as in the initial synthesis but the solid phase is notpooled and split for the final chemical coupling but are maintained asseparate pools then subjected to the chemical coupling step know forthat chapter. This resynthesis results in the formation of a new seriesof solid phase compound pools which have the last two chemical couplingsteps known. This new series of solid phase compound pools are screenedas in the initial screen and positive pools are checked as previouslyfor the binding specificity to identify positive pools. The positivepool(s) now allow the re-synthesis of the pool(s) with the last twosteps for the generation of the compound, which specifically binds tothe receptor subunit. The positive pools are then subjected to the samecycle of re-synthesis and screening as just described but with the lasttwo chemical coupling steps know the pools are maintained individuallyprior to the last know step. In this way the synthesis of the specificcompound able to bind to the receptor sub-unit is deconvoluted from thechemical library and identified.

In an alternative method the positive solid phase is removed from thescreen and collected. These are then subjected to the cleavage reaction,which removed the specific chemistry from the solid phase followed bythe analysis of the various chemical species using GC to separate theindividual compounds followed by MS to determine the molecular weight.This information coupled with the synthesis methods used is used todetermine the compound identity. After the determination of thesevarious candidate specific binding molecules they are thenre-synthesized and subjected to the binding assay to check if these arethe specific compounds that resulted in the positive solid phases.

Screening of the Ubiquitin Ligase Binding Moiety

This screening effort following methods and protocols known in the artallows the identification of compounds according to the presentinvention that bind to ubiquitin ligase.

These compounds, already identified here, form the basis for thedevelopment of compounds of the invention. These compounds are thensubjected to further chemistry based on the use of the linker group usedin the development of the solid phase chemistry. To this linker groupthe various ubiquitin ligase binding moieties and/or protein bindingmoieties are added, generally through condensation reactions or otherreactions to couple a ligand to a ubiquitin ligase binding moiety or aprotein binding moiety. These reactions are well known in the art.Derivatization of linker groups is well-known in the art and can consistof providing a nucleophilic group (e.g., an alcohol, amine, thiol orother nucleophilic group) or an electrophilic group (e.g., ester,carboxylic acid, acyl halide, halogen, etc.) at either or both ends ofthe linker group which may be used to condense an appropriately modifiedULM group and/or PTM group onto the linker to produce a covalent bond.This final step of chemistry generates the compounds of the invention.The compounds of the invention are then subject to analysis to determinewhich of the compounds from the chemical library screen with which ofthe individual ubiquitin ligase binding moiety element is able tofunction most effectively in the targeted ubiquitination and/ordegradation of the target protein. The ubiquitin ligase binding moietymay be determined by the methods as otherwise described in the examplessection hereinbelow. In addition, the compounds of the invention can betested in a mammalian tissue culture system where the target proteineither intact or as an engineered fragment is expressed. In such amammalian tissue culture system, the compounds effect on the targetprotein's level is determined by making use of the tag sequence whichcan be engineered into the recombinant expression of the target proteinduring the construction of the mammalian tissue culture test system. Thetag sequence is used to determine the levels of the target proteinduring the incubation with the potential compounds screened andsynthesized as described above. This assay for the tag sequence can takethe form of a western blot or via an ELISA, for example. Other tags,which are valuable to use are those based on the green fluorescentprotein, which allows the analysis of protein levels in living cellsand/or organisms.

The compounds that show the optimal activity in the test systems willthen form the basis for the next stage of drug development. In this nextstage these selected compounds are subjected to the recognized drugdevelopment path. The drug development path determines the potentialvalue of the compounds by evaluating a series of factors includingbioavailability; toxicology, pharmacology and efficacy in animal modelsbefore the compounds are considered for human testing.

Protein Level Control

This invention also relates to a method for the control of proteinlevels with a cell. This is based on the use of compounds of theinvention, which are known to interact with a specific target proteinsuch that degradation of a target protein in vivo will result in thecontrol of the amount of protein in a biological system, prerferably toa particular therapeutic benefit.

The following examples are used to assist in describing the presentinvention, but should not be seen as limiting the present invention inany way.

EXAMPLES First Set

The inventors initially hypothesized that small molecule inhibitors ofthe VHL/HIF-1α interaction could be rationally designed usinghydroxyproline (Hyp) as a starting point, since residue Hyp564 on HIF-1αmakes key interactions with VHL¹⁴ and is crucial for HIF-1α binding¹⁵.The inventors used the de-novo design software BOMB to guide theselection of plausible hydroxyproline analogs.¹⁶ 1 and 2 weresynthesized to test a promising design featuring an isoxazole moietypositioned to interact with a crystallographic water observed in thestructure of VHL bound to the HIF peptide (549-582)¹⁴ and a benzyl groupstacked along the side chain of Tyr98. Their ability to bind to VHL wasmeasured by the competition of a fluorescent HIF-1α peptide usingfluorescence polarization (FP).¹⁷ Both were able to displace thefluorescent peptide albeit at high concentrations (Table 1A). While thesmaller 3 was unable to fully displace the fluorescent peptide, theobserved binding to VHL through the use of WaterLOGSY and saturationtransfer difference (STD) NMR. As no binding was observed withhydroxyproline alone, this suggested that the inventors identified aminimal pharmacophore (see FIG. 2).

TABLE 1A Binding of Initial Ligands to VHL R IC₅₀ (μM)^(a) SEM (μM)  1 

  117   10   2

  120.1 7.1 3 CH₃ >250   N/A ^(a)Average IC₅₀ values were determinedfrom three independent trials, each in triplicate.

Encouraged by these initial results, the inventors sought to increasethe affinity of our VHL ligands by modifying the benzylamine moiety of 1while maintaining the methyl-isoxazole fragment. In order to generateanalogs rapidly, we developed a solid phase synthesis that involved theattachment of Fmoc-Hyp-OAllyl to Wang resin.¹⁸ Fmoc deprotection,coupling with 3-methyl-5-isoxazoleacetic acid followed by allyl esterdeprotection and coupling with various amines and subsequent cleavagewith trifluoroacetic acid led to the rapid generation of VHL ligands(Scheme 1).^(19,20) These ligands were then tested for their ability tobind VHL using the HIF peptide FP displacement assay.

Incorporation of various halogenated benzylamines showed that parasubstitution yielded the highest affinity and that there were onlyslight differences of affinity between chlorides and bromides, althoughthe corresponding fluoride was less potent. We also found thatsubstitution with electron withdrawing groups such as esters, nitrogroups, nitriles, and ketones led to more potent ligands thansubstitution with the electron donating methoxy and t-butylsubstituents. Molecular dynamics simulations suggested that Arg107 isflexible and could accommodate bulkier groups at the para position.Therefore, we considered larger heterocyclic substituents at the paraposition of the benzylamine moiety and synthesized 15, which was foundto bind with a 4.1 μM IC₅₀ value (Table 2, below).

Fluorescence Polarization Assay

Ability of VHL ligands to compete for the HIF 1α binding site on VCB wasdetermined through a fluorescence polarization competition assay asdescribed in the literature (Buckley et al. JACS, 2012, 134, 4465-4468).VHL ligands were dissolved in DMSO (100 mM), and then diluted 10 foldwith VHL buffer. The compounds were then diluted 2 fold with 10% DMSO inbuffer (20 uL into uL of 10% DMSO in buffer) 14 times. AqueousDEALA-Hyp-YIPD was used as a positive control. 278 nM FAM-DEALA-Hyp-YIPD(DMSO) was diluted 1000 fold into VHL buffer. For polarizationdisplacement assays, 9 uL of 1 uM V1-213CB (450 nM final), 2 uL of VHLLigands (VL) compounds in 10% DMSO (1% DMSO final), and 9 uL of 278 nMFAM-DEALA-Hyp-YIPD were added to a 384 well plate (Corning 3575). Theplate was then shaken for 1 minute, and centrifuged for 1 minute, beforereading fluorescence polarization on a Perkin Elmer Envision 2101Multilabel reader (excitation 486 nM, emission 535 nM). Wells containingV1-213CB, DMSO vehicle, FAM-DEALA-Hyp-YIPD served as maximumpolarization (or minimum displacement). Wells containing buffer in placeof V1-213CB, DMSO vehicle, FAM-DEALA-Hyp-YIPD served as minimumpolarization (or maximum displacement). The percent inhibition wasdetermined by normalization to maximum and minimum polarization, andgraphed against the log [VL]. IC50 values were then determined usingPrism 5 for each replicate (n=9), which were then averaged to determinethe average IC50 and the standard error of the mean (SEM). The resultsfor a number of exemplary compounds are presented in Table 2 AffinityTable herein below.

TABLE 2 Affinity Table. VHL IC₅₀ (μM) IC₅₀ (μM) ligand No. 10% DMSO 1%DMSO Chemical Structure VL001 >1000

VL002 241.1 120

VL003 >1000

VL004 123.1 117

VL005 >1000

VL006 Inactive

VL007 Inactive

VL008 >1000

VL009 Inactive

VL010 Inactive

VL011 >1000

VL012 >1000

VL013 240

VL014 4.7

VL015 >1000

VL016 ~730

VL017 ~510

VL018 280

VL019 450.5

VL020 295.4 149

VL021 36 20.5

VL022 ~870

VL023 589

VL024 511

VL025 517

VL026 >1000 >250

VL027 >1000

VL028 210.0 149

VL029 24.9 32

VL030 215

VL031 68.5 >250

VL032 639

VL033 >1000

VL034 130.7 130

VL035 34 39.4

VL036 743.5

VL037 88.8

VL039 >1000

VL043 >1000

VL044 285.8

VL045 145.6

VL046 215.2

VL047 405.7 106

VL048 24.8 16

VL049 15.1

VL050 332.1

VL051 >1000

VL052 >1000

VL053 >1000

VL054 139.6

VL055 910.3

VL056 938.5

VL057 >1000

VL058 >1000

VL059 205.4

VL060 >1000

VL061 419.5

VL062 >1000

VL063 >1000

VL064 110.0

VL065 74.2

VL066 72.4

VL067 70.7

VL068 >1000

VL069 23.8

VL070 38.2

VL071 10.4

VL072 29.5

VL073 39 46

VL074 42.9

VL075 35.5

VL076 19.6

VL077 8.9

VL078 60.2

VL079 26.3

VL080 17.0

VL081 15.5

VL082 29.0

VL083 >1000

VL084 460.3

VL085 27.1

VL086 168.4

VL087 102.6

VL088 18.2 60

VL089 40.8

VL090 45.2

VL091 23.0

VL093 510

VL094 540

VL095 14.8 22.6

VL096 32.5

VL097 49.4

VL098 244.4

VL099 39.6

VL100 63.6

VL101 940

VL102 64.5

VL104 >1000

VL105 >1000

VL106 >1000

VL108 347.2

VL109 349.0

VL110 138.0

VL111 4.1 4.1

VL112 11.3

VL113 Inactive Inactive

VL114 Inactive Inactive

VL115 17.0 19

VL116 2.9 2.9

VL117 5.0 13

VL118 16.4 31

VL119 17.8 33

VL120 >1000 >1000

VL121 >1000 >1000

VL122 102.1 220

VL123 560 404

VL124 990 670

VL125 790 568

VL126 88.1 82

VL127 47.7 73

VL128 57.9 224

VL129 760 493

VL130 >1000 >1000

VL131 8.0 19

VL132 77.7 85

VL133 2.2 7.5

VL134 48.1 54

VL135 Inactive >1000

VL136 28.2 540

VL137 35.6 128

VL138 52.2 70

VL139 24.7 44

VL140 10.3 17

VL141 191.8 220

VL142 780 >1000

VL143 >1000 >1000

VL144 >1000 >1000

VL145 >1000 >1000

VL146 128.9 196

VL147 >1000 >1000

VL148 23.8 113

VL149

VL150 16.0 41

VL151 11 33

VL152 22.8 32

VL153 978.3

VL154 118.3 77

VL155 17.4 14

VL156 >1000 >1000

VL157 371.7 360

VL158 265.8 180

VL159 66.9 54

VL160 12.1 9

VL161 36.4 35

VL162 235.4 250

VL163 507.5 610

VL164 72.9 40

VL165

VL166

VL167

VL168

VL169

VL170

VL171

VL172

VL173

VL174

VL175

VL176

VL177

VL178

VL179

VL180

VL181

VL182

VL183

VL184

VL185

VL186

VL187

VL188

VL189

VL190

VL191

VL192

VL193

VL194

VL195

VL196

VL197

VL198

VL199

VL200

VL201

VL202

VL203

VL204

VL205

VL206

VL207

VL208

VL209

VL210

VL211

VL212

VL213

VL214

VL215

VL216

VL217

VL218

VL219

VL220

VL221

VL222

VL223

VL224

VL225

VL226

VL227

VL228

VL229

VL230

VL231

VL232

VL237

VL238

VL239

VL240

VL241

VL242

VL243

VL244

VL245

VL247

VL248

VL249

VL250

VL251

VL252

VL253

VL254

VL255

VL256

VL257

VL258

VL259

VL260

VL261

VL262

VL263

VL264

VL265

VL266

Synthetic Methods General Chemistry

All reactions were performed in oven-dried or flame-dried glasswarefitted with rubber septa under a positive pressure of nitrogen, unlessotherwise noted. Air- and moisture-sensitive liquids were transferredvia syringe or cannula. THF was distilled from sodium/benzophenone.Dichloromethane was distilled from calcium hydride. Analytical thinlayer chromatography (TLC) was performed using glass plates precoatedwith silica gel (0.25 mm). TLC plates were visualized by exposure to UVlight (UV) or KMnO₄. Flash column chromatography was performed usingsilica gel 60 (230-400 mesh, Merck) with the indicated solvents.

¹H and ¹³C spectra were recorded on Bruker Avance DPX-500 or BrukerAvance DPX-400 NMR spectrometers. ¹H NMR spectra are represented asfollows: chemical shift, multiplicity (s=singlet, d=doublet, t=triplet,q=quartet, m=multiplet, br=broad), integration, and coupling constant(J) in Hertz (Hz). ¹H NMR chemical shifts are reported relative to CDCl₃(7.26 ppm), d₆-DMSO (2.50 ppm) and d₄-MeOD (3.31 ppm). ¹³C NMR wasrecorded relative to the central line of CDCl₃ (77.16 ppm), d₆-DMSO(39.52 ppm) and d₄-MeOD (49.00 ppm). In most cases, only peaks of themajor rotamer are reported. Mass spectra were obtained using aPerkin-Elmer API 150 EX spectrometer. MALDI-TOF analyses of purifiedsamples were performed in a Voyager-DE-PRO 6268 (Applied Biosystems)using cyano-4-hydroxycinnamic acid matrices. Unless otherwise noted,HPLC was performed using a Dynamax SD200 solvent delivery systemconnected to a Dynamax UV-1 Absorbance Detector with a YMC-Pack ODS-AMpreparative column (250×20 mm, 5 m particle size, 12 nm pore size). Alinear gradient of MeCN in H₂O from 20% to 100% MeCN, with constant 0.1%TFA was run over 40 minutes.

General Methods of Chemical Synthesis

The following eight (8) general chemical synthetic methods (Methods Athrough F and Solid Phase Synthesis A and B, described hereinbelow) areprovided for synthesizing numerous compounds according to the presentinvention which are set forth in Table 2 Affinity Table above. Eachmethod is presented with reference to a specific compound, the syntheticdetails of which are presented hereinabove. All of the compoundsnumbered may be synthesized relatively easily using the straight-forwardmethods which are set forth hereinbelow. In certain instances, moresynthetic details are provided for certain preferred embodiments inorder to present that information such that it may serve as a templatefor synthesizing a number of other compounds as otherwise disclosedherein.

As an example, see the synthesis for compound VL133 of Table II, setforth below.

See the general synthesis for VL116 of Table II with protection of thehydroxyl group.

See the general synthesis for compound VL 156 of Table II, describedbelow.

See the general synthesis for compound VL 217 of Table II, describedbelow.

See the general synthesis for VL 219 of Table II, described below.

Method F subsumes methods C, D and E and is a general method whichproceeds through commercially available amines.

Following the general synthetic methods set forth above and aspreviously described, the following compounds are synthesized byanalogy.

(2S,4R)-1-((9H-fluoren-9-yl)methyl) 2-allyl4-(tert-butoxy)pyrrolidine-1,2-dicarboxylate (Fmoc-Hyp(OtBu)-OAllyl)

Fmoc-Hyp(OtBu)OH (24.9 g, 60.8 mmol, 1 eq) was dissolved in DMF (300 mL)at room temperature. Sodium bicarbonate (12.8 g, 152 mmol, 2.5 eq) wasadded, followed by allyl bromide (25.3 mL, 300 mmol, 4.9 eq). Thesolution was fitted with an air condenser and heated to 50° C. for 20hours. It was then cooled to room temperature, diluted with EtOAc,washed with aqueous 1 M HCl, saturated sodium bicarbonate, water andbrine. The organic layer was dried with sodium sulfate, filtered andcondensed.¹⁵ Purification by column chromatography (15 to 33%EtOAc/hexanes) gave Fmoc-Hyp(OtBu)OAllyl (23.42 g, 52.1 mmol, 86%) as afaint yellow oil. ¹H NMR (500 MHz, CDCl₃) δ 7.76 (t, J=6.3 Hz, 2H),7.63-7.54 (m, 2H), 7.43-7.37 (m, 2H), 7.31 (t, J=7.0 Hz, 2H), 5.99-5.79(m, 1H), 5.39-5.18 (m, 2H), 4.66 (d, J=5.6 Hz, 1H), 4.63-4.13 (m, 6H),3.81 (ddd, J=16.6, 10.7, 6.2 Hz, 1H), 3.48-3.33 (m, 1H), 2.31-2.18 (m,1H), 2.18-2.08 (m, 1H), 1.21 (d, J=11.6 Hz, 9H). ¹³C NMR (126 MHz,CDCl₃) (mixture of rotamers) δ 172.49, 155.01, 154.49, 144.31, 144.18,144.06, 143.84, 141.44, 141.41, 141.36, 131.91, 131.74, 127.80, 127.76,127.20, 127.16, 125.31, 125.28, 125.11, 120.08, 120.05, 118.93, 118.61,74.29, 69.37, 68.48, 67.73, 65.86, 58.09, 57.79, 54.01, 53.52, 47.40,47.28, 38.90, 37.87, 28.41, 28.37. MS (ESI) 450.5 (M+H).

(2S,4R)-1-((9H-fluoren-9-yl)methyl) 2-allyl4-hydroxypyrrolidine-1,2-dicarboxylate (Fmoc-Hyp(OH)-OAllyl)

Fmoc-Hyp(OtBu)-OAllyl (23.42 g, 52.1 mmol) was dissolved in DCM (306 mL)at room temperature. TFA (54 mL, 15% vol/vol) was added and the solutionwas stirred for 13 hours. The solution was poured into water,neutralized by slow addition of saturated aqueous sodium bicarbonate andextracted twice with DCM and once with EtOAc. The combined organiclayers were dried with sodium sulfate, filtered and condensed.Purification by column chromatography (30 to 80% EtOAc/hexanes) gaveFmoc-Hyp(OH)-OAllyl as a yellowish oil (16.7 g, 42.4 mmol, 81%). ¹H and¹³C NMR spectra matched those reported in the literature.¹⁶

Fmoc-Hyp(OWang)-OAllyl

Wang Resin (12.1 g, 1.1 mmol/g loading, 13.3 mmol, 1 eq) was swelledwith DCM (90 mL) in a glass reaction vessel and cooled to 4° C.Trichloroacetonitirle (20 mL, 200 mmol, 15 eq) was added, followed bythe addition of DBU (3 mL, 20 mmol, 1.5 eq) in 3 portions over 3minutes, manually shaking the reaction vessel in between additions. Thereaction vessel was nutated at 4° C. for 1 hour, then washed with DCM,DMSO, THF, then twice with DCM at room temperature.¹⁷ A solution ofFmoc-Hyp(OH)-OAllyl (26.15 g, 66.5 mmol, 5 eq) in DCM (40 mL) and THF(40 mL) was then added, and shaken for 30 minutes and then washed twicewith DCM, thrice with DCM and then twice with MeOH followed by DCM. Theinitial DCM washes were condensed, and purified by column chromatography(33% to 80% EtOAc) to recover the Fmoc-Hyp(OH)-OAllyl starting material(21.51 g, 54.67 mmol, 82%). The resin was dried in air, then dried undervacuum to give 15.5 g of Fmoc-Hyp(OWang)-OAllyl.¹⁸ The loading of theresin was estimated to be 0.53 mmol/g based upon the increase in mass.

Solid Phase Synthesis General Method A

Fmoc-Hyp(OWang)-OAllyl resin (1 eq) was swelled DMF, then reacted with20% piperidine in DMF for 30 minutes. The resin was then washed oncewith piperidine, and reacted again with 20% piperidine for 30 minutes toensure complete deprotection. The resin was then washed twice with DMFand once with MeOH followed by DCM. The resulting free amine was thencoupled with 3-methyl-5-isoxazoleacetic acid (4 eq), PyBOP (4 eq) HOBt(4 eq) and DIPEA (7 eq) in DMF for 4 hours. The resin was then washedthrice with DMF and twice with MeOH followed by DCM. The resin was thenswelled with freshly distilled DCM, and reacted with Pd(PPh₃)₄(0.1 eq)and PhSiH₃ (10 eq) for 30 minutes. The resin was then washed once withDCM, and reacted again with Pd(PPh₃)₄(0.1 eq) and PhSiH₃ (10 eq) indistilled DCM for 30 minutes, after which the resin was washed twicewith DMF and once with MeOH followed by DCM. The resulting carboxylicacid was then coupled with the appropriate amine (or a salt of theappropriate amine), RNH₂ (4 eq) with PyBOP (4 eq), HOBt (4 eq) and DIPEA(7 eq for free amines, 8 eq for amine salts) in DMF for 4 hours. Theresin was then washed 5 times with DMF, thrice with MeOH and 5 timeswith DCM. The resin was then reacted with 20% TFA in DCM for 2 hours.The reaction mixture was then drained and the resin was washed with DCM.Condensation under reduced pressure, and purification by columnchromatography (1% to 10% 0.5M NH₃ in MeOH/DCM or 1% to 10% MeOH in DCM)gave the desired VHL ligand.

Solid Phase Synthesis General Method B

Briefly, Fmoc-Hyp-(OWang)-OAllyl resin (1 eq) was swelled with freshlydistilled DCM, and reacted with Pd(PPh₃)₄(0.1 eq) and PhSiH₃ (10 eq) for30 minutes. The resin was then washed once with DCM, and reacted againwith Pd(PPh₃)₄(0.1 eq) and PhSiH₃ (10 eq) in distilled DCM for 30minutes, after which the resin was washed twice with DMF and once withMeOH followed by DCM. The resulting carboxylic acid was then coupledwith 4-chlorobenzylamine (4 eq), PyBOP (4 eq), HOBt (4 eq) and DIPEA (7eq) in DMF for 4 hours. The resin was then reacted with 20% piperidinein DMF for 30 minutes. The resin was then washed once with DMF, andreacted again with 20% piperidine for 30 minutes to ensure completedeprotection. The resin was then coupled with the appropriate carboxylicacid (RCO₂H, 4 eq), PyBOP (4 eq), HOBt (4 eq) and DIPEA (7 eq) in DMFfor 4 hours. The resin was then washed 4 times with DMF and twice withmethanol followed by DCM. The resin was then reacted with 20% TFA in DCMfor 2 hours. The reaction mixture was then drained and the resin waswashed with DCM. Condensation under reduced pressure, and purificationby column chromatography (1% to 10% 0.5M NH₃ in MeOH/DCM or 1% to 10%MeOH in DCM) gave the desired VHL ligand. Yields are based upon theloading of the resin, which was estimated based upon its change in mass.

tert-Butyl 4-(methoxy(methyl)carbamoyl)benzylcarbamate(Boc-Amb-N(OMe)Me)

Boc-Amb-OH (2.55 g, 10.16 mmol, 1 eq) was dissolved in DCM (68 mL) andcooled to 4° C. in an ice bath. EDC (2.34 g, 12.2 mmol, 1.2 eq), HOBt(1.65 g, 12.2 mmol, 1.2 eq) and DIPEA (6.2 mL, 35.6 mmol, 3.5 eq) wereadded. The solution was stirred for 30 minutes and thenN,O-Dimethylhydroxylamine hydrochloride (1.09 g, 11.2 mmol, 1.1 eq) wasadded. The solution warmed slowly to room temperature and after 21 hourswas poured into brine, with a small amount of chloroform to break theresulting emulsion. After separation, the aqueous layer was extractedtwice with EtOAc. The combined organic layer was dried over sodiumsulfate, filtered and condensed. Purification by column chromatography(40 to 75% EtOAc/hexanes) gave a colorless oil (2.45 g, 8.33 mmol, 82%).¹H NMR (500 MHz, CDCl₃) δ 7.65 (d, J=8.2 Hz, 2H), 7.31 (d, J=8.1 Hz,2H), 4.88 (s, 1H), 4.36 (d, J=5.1 Hz, 2H), 3.55 (s, 3H), 3.35 (d, J=4.7Hz, 3H), 1.47 (s, 9H). ¹³C NMR (126 MHz, CDCl3) δ 169.77, 156.04,141.79, 133.21, 128.76, 127.03, 79.87, 61.20, 44.50, 33.88, 28.55. MS(ESI) 295.2 (M+H).

tert-Butyl 4-formylbenzylcarbamate (Boc-Amb-H)

Boc-Amb-N(OMe)Me (2.45 g, 8.33 mmol, 1 eq) was dissolved in THF (83 mL)and cooled to −78° C. in a dry ice/acetone bath. Lithium aluminumhydride (0.41 g, 10.83 mmol, 1.3 eq) was added in 2 portions over 5minutes. After 50 minutes, the suspension was warmed to 4° C. in an icebath. After 3.5 hours, the reaction was deemed complete by TLC (miniworkup in 10% potassium bisulfate and EtOAc, 50% EtOAc/hexanes) and thereaction was quenched by the slow addition of 10% potassium bisulfate at4° C. The mixture was warmed to room temperature, and stirred for 30minutes. Most of the THF was removed under reduced pressure and mixturewas diluted with water and extracted thrice with EtOAc. The combinedorganic layer was washed once with brine, dried over sodium sulfate,filtered and condensed. Purification by column chromatography (40 to 50%EtOAc/hexanes) gave Boc-Amb-H as a white solid (1.66 g, 7.1 mmol, 85%).¹H NMR (500 MHz, CDCl₃) δ 9.96 (s, 1H), 7.81 (d, J=8.2 Hz, 2H), 7.41 (d,J=8.0 Hz, 2H), 5.12 (s, 1H), 4.37 (d, J=5.6 Hz, 2H), 1.44 (s, 9H). ¹³CNMR (126 MHz, CDCl₃) δ 191.94, 156.03, 146.30, 135.62, 130.14, 127.78,79.92, 44.44, 28.46. MS (ESI) 235.9 (M+H), 180.2 (M-tBu).

tert-Butyl 4-(oxazol-5-yl)benzylcarbamate

Potassium carbonate (0.13 g, 0.94 mmol, 1.2 eq) andtoluenesulfonylmethyl isocyanide (0.184 g, 0.94 mmol, 1.2 eq) were addedto MeOH (7.8 mL) at room temperature. The round bottom was fitted with areflux condenser and heated to 45° C. After 15 minutes, Boc-Amb-H(0.1835 g, 0.78 mmol, 1 eq) was added and the mixture was heated to 75°C. for 3 hours and then cooled to room temperature. The MeOH was removedunder reduced pressure and the crude material was resuspended in EtOAcand 1:2 mixture of saturated sodium carbonate to water and separated.The aqueous layer was then extracted once with EtOAc. The combinedorganic layer was dried over sodium sulfate, filtered and condensed.Purification by column chromatography (20 to 35% EtOAc/hexanes) gave awhite solid. ¹H NMR (400 MHz, CDCl₃) δ 7.91 (s, 1H), 7.62 (d, J=8.3 Hz,2H), 7.35 (ob d, 2H), 7.34 (ob s, 1), 4.88 (s, 1H), 1.47 (s, 9H). ¹³CNMR (126 MHz, CDCl₃) δ 156.02, 151.40, 150.47, 139.78, 128.01, 126.84,124.67, 121.47, 79.68, 44.38, 28.47. MS (ESI) 275.5 (M+H).

(4-(Oxazol-5-yl)phenyl)methanamine trifluoroacetate salt

To a solution of tert-butyl 4-(oxazol-5-yl)benzylcarbamate (1.09 g) inDCM (40 mL), TFA (4 mL) was added at room temperature. The solution wasstirred for 16 hours and concentrated under reduced pressure to yieldthe trifluoroacetate salt of (4-(oxazol-5-yl)phenyl)methanamine (1.984g) as a cream colored solid, which was used without furtherpurification. ¹H NMR (400 MHz, MeOD) δ 8.29 (s, 1H), 7.83 (d, J=8.4 Hz,2H), 7.60 (s, 1H), 7.56 (d, J=8.5 Hz, 2H), 4.16 (s, 2H). MS (ESI) 175.3(M-CF₃CO₂ ⁻).

(2S,4R)—N-(3-chlorobenzyl)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide(VL4)

VL4 was synthesized according to General Method F as a white solid. ¹HNMR (500 MHz, MeOD): δ8.684 (1H, s); 7.33-7.23 (4H, m); 6.24 (1H, s);4.56-4.53 (1H, t, J=8 Hz); 4.51-4.50 (1H, m); 4.39-4.37 (2H, m);3.96-3.92 (2H, m); 3.81-3.3.78 (1H, dd, J=9 Hz, 4 Hz); 3.64-3.62 (1H,m); 2.28-2.24 (4H, m); 2.09-2.04 (1H, m).¹³C NMR (125 MHz, MeOD):δ174.56, 168.67, 167.68, 161.58, 142.25, 135.35, 131.04, 128.43, 128.19,126.76, 105.37, 70.86, 60.78, 56.96, 43.60, 39.33, 33.90, 11.21. MS(ESI) 378.2 (M+H).

(2S,4R)-4-hydroxy-N-(4-hydroxyphenethyl)-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide(VL2)

VL2 was synthesized according to General Method F. ¹H NMR (500 MHz,MeOD) δ 8.33 & 8.13 (due to the rotamers, both s, 1H), 7.02 (d, J=8.3Hz, 2H), 6.69 & 6.65 (due to the rotamers, both d, J=8.3 Hz, 2H), 6.22 &6.10 (due to the rotamers, both s, 1H), 4.43 (d, J=7.7 Hz, 2H), 3.89 (d,J=4.7 Hz, 2H), 3.74 (dd, J=11.0, 4.3 Hz, 1H), 3.57 (d, J=11.0 Hz, 1H),3.42-3.36 (m, 2H), 2.73-2.63 (m, 2H), 2.25 (s, 3H), 2.16-2.12 (m, 1H),1.96-1.91 (m, 1H). ¹³C NMR (asterisk denotes the signals of the minorrotamer, 125 MHz, MeOD) δ 174.2, 174.1, *173.9, *173.8, *169.0, 168.6,167.6, *167.4, 161.6, *161.5, *157.0, 156.9, *131.2, 130.8, *116.2,116.1, *105.6, 105.4, 70.7, *69.2, *61.0, *60.9, 60.7, 60.6, 56.9,*56.2, 42.4, 42.3, *42.0, *41.9, 41.5, 39.3, 35.5, *35.3, 33.9, *32.9,11.2. MS (ESI) [M+H] 374.1, [2M+Na] 769.6.

(2S,4R)-4-hydroxy-N-methyl-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide(VL26)

VL26 was synthesized according to General Method F and was isolated as acolorless oil (28 mg, 0.105 mmol, 80%). ¹H NMR (500 MHz, MeOD) δ 6.23(s, 1H), 4.47 (dt, J=16.3, 5.2 Hz, 2H), 3.91 (d, J=5.7 Hz, 2H), 3.78(dd, J=10.9, 4.2 Hz, 1H), 3.61 (dd, J=11.0, 1.8 Hz, 1H), 2.73 (s, 3H),2.26-2.18 (m, 4H), 2.05 (dd, J=8.3, 4.7 Hz, 1H). ¹³C NMR (126 MHz, MeOD)δ 174.82, 168.66, 167.67, 161.58, 105.40, 70.79, 60.67, 56.91, 39.30,33.89, 26.35, 11.20. MS (ESI) 291.1 (M+Na), 268.7 (M+H).

VL34 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.3 mmol) according toSolid Phase Synthesis General Method A. It was isolated as a white solid(14.7 mg). ¹H NMR (400 MHz, MeOD) δ 7.31 (dd, J=5.9, 5.1 Hz, 4H),7.27-7.17 (m, 1H), 6.23 (s, 1H), 4.55 (t, J=8.0 Hz, 1H), 4.50 (s, 1H),4.39 (s, 2H), 3.92 (d, J=1.8 Hz, 2H), 3.80 (dd, J=10.9, 4.3 Hz, 1H),3.61 (dd, J=7.3, 5.5 Hz, 1H), 2.33-2.19 (m, 4H), 2.12-2.03 (m, 1H). ¹³CNMR (101 MHz, MeOD) δ 174.29, 168.68, 167.68, 161.60, 139.73, 129.51,128.40, 128.14, 105.36, 70.84, 60.73, 56.97, 44.05, 39.36, 33.95, 11.21.MS (ESI) 344.3 (M+H), 366.2 (M+Na).

VL28 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.3 mmol) according toSolid Phase Synthesis General Method A. It was isolated as a yellowsolid (19.1 mg). ¹H NMR (500 MHz, MeOD) δ 8.66 (t, J=5.5 Hz, 1H),7.48-7.34 (m, 2H), 7.31-7.21 (m, 2H), 6.23 (s, 1H), 4.58 (t, J=8.0 Hz,1H), 4.48 (qd, J=15.8, 5.9 Hz, 3H), 3.99-3.87 (m, 2H), 3.80 (dd, J=10.9,4.3 Hz, 1H), 3.66-3.60 (m, 1H), 2.31-2.22 (m, 4H), 2.09 (ddd, J=13.0,8.2, 4.7 Hz, 1H). ¹³C NMR (126 MHz, MeOD) δ 174.60, 168.72, 167.65,161.59, 136.79, 134.01, 130.29, 130.08, 129.67, 128.21, 105.37, 70.84,60.74, 56.96, 42.08, 39.34, 33.95, 11.22. MS (ESI) 378.3 (M+H).

VL21 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.2 mmol) using SolidPhase Synthesis General Method A. It was isolated as a white solid (15.9mg). ¹H NMR (500 MHz, MeOD) δ 8.65 (s, 1H), 7.32-7.26 (m, 4H), 6.22 (s,1H), 4.58-4.47 (m, 2H), 4.43-4.32 (m, 2H), 3.92 (d, J=4.2 Hz, 2H), 3.80(dd, J=10.9, 4.3 Hz, 1H), 3.66-3.58 (m, 1H), 2.30-2.22 (m, 4H), 2.08(dd, J=8.3, 4.7 Hz, 1H). ¹³C NMR (126 MHz, MeOD) δ 174.48, 168.71,167.66, 161.60, 138.67, 133.85, 129.99, 129.53, 105.37, 70.85, 60.80,56.99, 43.45, 39.33, 33.95, 11.20. MS (ESI) 378.4 (M+H).

VL20 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.15 mmol) using SolidPhase Synthesis General Method A. It was isolated as a white solid (9.9mg). ¹H NMR (400 MHz, MeOD) δ 8.64 (t, J=5.6 Hz, 1H), 7.37-7.26 (m, 2H),7.07-6.99 (m, 2H), 6.23 (s, 1H), 4.57-4.47 (m, 2H), 4.37 (dd, J=8.4, 5.8Hz, 2H), 3.93 (d, J=3.0 Hz, 2H), 3.80 (dd, J=11.0, 4.2 Hz, 1H),3.66-3.58 (m, 1H), 2.28-2.22 (m, 4H), 2.08 (dd, J=8.3, 4.7 Hz, 1H). ¹³CNMR (126 MHz, MeOD) δ 174.31, 168.69, 167.67, 163.44 (d, J=243.5 Hz),161.60, 135.78, 130.27 (d, J=8.1 Hz), 116.09 (d, J=21.6 Hz), 105.37,70.85, 60.75, 56.99, 43.32, 39.33, 33.95, 11.20. MS (ESI) 362.3 (M+H).

VL29 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.3 mmol) according toSolid Phase Synthesis General Method A. It was isolated as a lightyellow solid (16.4 mg). ¹H NMR (400 MHz, MeOD) δ 7.45 (dq, J=9.0, 2.2Hz, 2H), 7.23 (d, J=8.5 Hz, 2H), 6.22 (s, 1H), 4.58-4.47 (m, 2H), 4.35(dt, J=18.9, 15.4 Hz, 2H), 3.92 (d, J=2.6 Hz, 2H), 3.80 (dd, J=10.9, 4.2Hz, 1H), 3.63 (d, J=11.0 Hz, 1H), 2.30-2.21 (m, 4H), 2.11-2.02 (m, 1H).¹³C NMR (101 MHz, MeOD) δ 174.41, 168.71, 167.66, 161.61, 139.15,132.55, 130.32, 121.77, 105.37, 70.85, 60.75, 56.99, 43.37, 39.33,33.94, 11.22. MS (ESI) 424.1 (M+H).

VL31 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.3 mmol) according toSolid Phase Synthesis General Method A. It was isolated as a white solid(19.8 mg). ¹H NMR (500 MHz, MeOD) δ 8.56 (s, 1H), 7.35 (d, J=8.2 Hz,2H), 7.22 (d, J=8.2 Hz, 2H), 6.24 (s, 1H), 4.53 (dd, J=18.3, 10.3 Hz,2H), 4.36 (d, J=5.7 Hz, 2H), 3.92 (d, J=3.0 Hz, 2H), 3.80 (dd, J=10.9,4.2 Hz, 1H), 3.62 (d, J=11.1 Hz, 1H), 2.29-2.21 (m, 4H), 2.12-2.02 (m,1H), 1.29 (d, J=7.9 Hz, 9H). ¹³C NMR (126 MHz, MeOD) δ 174.29, 168.67,167.68, 161.59, 151.18, 136.67, 128.19, 126.39, 105.38, 70.83, 60.77,56.97, 43.88, 39.37, 35.28, 33.95, 31.79, 31.74, 11.23. MS (ESI) 400.5(M+H).

VL47 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.156 mmol) accordingto Solid Phase Synthesis General Method A. It was isolated as a whitesolid (9.1 mg). ¹H NMR (500 MHz, MeOD) δ 7.22 (dd, J=8.4, 3.9 Hz, 2H),6.86 (dd, J=8.8, 2.2 Hz, 2H), 6.22 (s, 1H), 4.63-4.45 (m, 2H), 4.37-4.26(m, 2H), 3.92 (d, J=2.6 Hz, 2H), 3.83-3.70 (m, 4H), 3.61 (d, J=11.2 Hz,1H), 2.28-2.20 (m, 4H), 2.06 (ddd, J=13.0, 8.1, 4.7 Hz, 1H). ¹³C NMR(126 MHz, MeOD) δ 174.13, 168.66, 167.68, 161.60, 160.39, 131.67,129.76, 114.89, 105.37, 70.83, 60.74, 56.97, 55.67, 43.57, 39.34, 33.95,11.21. MS (ESI) 374.5 (M+H).

VL35 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.156 mmol) accordingto Solid Phase Synthesis General Method A. It was isolated as a whitesolid (14.1 mg). ¹H NMR (500 MHz, DMSO) δ 7.90-7.85 (m, 2H), 7.39 (d,J=8.4 Hz, 2H), 6.23 (s, 1H), 5.17 (s, 1H), 4.37 (dd, J=17.9, 10.4 Hz,4H), 3.88 (s, 2H), 3.84 (s, 3H), 3.70 (dd, J=10.5, 4.6 Hz, 1H), 3.47(dd, J=10.4, 2.5 Hz, 1H), 2.20 (d, J=10.2 Hz, 3H), 2.11-2.03 (m, 1H),1.92 (ddd, J=12.5, 7.2, 4.9 Hz, 1H). ¹³C NMR (126 MHz, DMSO) δ 171.66,166.66, 166.10, 165.66, 159.35, 145.22, 129.08, 127.99, 127.06, 103.94,68.62, 58.76, 55.20, 52.02, 41.49, 38.17, 32.73, 10.95. MS (ESI) 402.6(M+H).

VL48 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.156 mmol) accordingto Solid Phase Synthesis General Method A. It was isolated as a whitesolid (11.4 mg). ¹H NMR (400 MHz, MeOD) δ 8.28-8.05 (m, 2H), 7.55 (d,J=8.8 Hz, 2H), 6.23 (s, 1H), 4.64-4.36 (m, 4H), 3.94 (d, J=3.8 Hz, 2H),3.81 (dd, J=10.9, 4.2 Hz, 1H), 3.65 (dt, J=11.0, 1.7 Hz, 1H), 2.34-2.21(m, 4H), 2.09 (td, J=8.5, 4.2 Hz, 1H). ¹³C NMR (101 MHz, MeOD) δ 174.70,168.79, 167.65, 161.63, 148.48, 147.72, 129.13, 124.56, 105.40, 70.88,60.79, 57.03, 43.41, 39.32, 33.94, 11.20. MS (ESI) 389.3 (M+H), 411.4(M+Na).

VL88 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.156 mmol) accordingto Solid Phase Synthesis General Method A. It was isolated as a clearoil (8.0 mg). 1H NMR (500 MHz, MeOD) δ 8.77 (s, 1H), 7.58 (dd, J=88.0,8.1 Hz, 4H), 6.23 (d, J=4.4 Hz, 1H), 4.61-4.33 (m, 4H), 3.93 (d, J=9.7Hz, 2H), 3.83-3.74 (m, 1H), 3.63 (dd, J=10.4, 9.0 Hz, 1H), 2.33-2.27 (m,1H), 2.26 (d, J=3.7 Hz, 3H), 2.15-2.03 (m, 1H). ¹³C NMR (126 MHz, MeOD)δ 175.33, 168.76, 167.64, 161.61, 145.83, 133.39, 129.12, 119.74,111.79, 105.28, 70.87, 70.87, 59.32, 57.02, 43.61, 38.79, 33.94, 11.21,11.19. MS (ESI) 391.2 (M+Na).

VL95 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.156 mmol) accordingto Solid Phase Synthesis General Method A. It was isolated as a whitesolid (23 mg). 1H NMR (400 MHz, MeOD) δ 8.71 (s, 1H), 7.96-7.91 (m, 2H),7.43 (d, J=8.5 Hz, 2H), 6.22 (s, 1H), 4.56 (t, J=8.0 Hz, 1H), 4.49 (ddd,J=18.6, 8.6, 4.1 Hz, 3H), 3.91 (s, 2H), 3.80 (dd, J=10.9, 4.2 Hz, 1H),3.65-3.58 (m, 1H), 2.58 (d, J=1.6 Hz, 3H), 2.28-2.22 (m, 4H), 2.10 (dd,J=8.3, 4.7 Hz, 1H). ¹³C NMR (101 MHz, MeOD) δ 200.08, 174.34, 168.43,167.36, 161.39, 145.50, 136.96, 129.62, 128.28, 105.26, 70.65, 60.57,56.83, 43.72, 39.14, 33.87, 26.73, 11.30. MS (ESI) 386.0 (M+H).

VL111 was synthesized from Fmoc-Hyp(OWang)-OAllyl (0.2 mmol) accordingto Solid Phase Synthesis General Method A. It was isolated as a whitesolid (18.2 mg). ¹H NMR (500 MHz, MeOD) δ 8.23 (s, 1H), 7.68 (d, J=8.1Hz, 2H), 7.49 (s, 1H), 7.41 (d, J=8.1 Hz, 2H), 6.23 (s, 1H), 4.59-4.37(m, 4H), 3.93 (d, J=3.4 Hz, 2H), 3.81 (dd, J=10.9, 4.1 Hz, 1H), 3.63 (d,J=11.0 Hz, 1H), 2.32-2.17 (m, 4H), 2.09 (ddd, J=13.0, 8.0, 4.6 Hz, 1H).¹³C NMR (126 MHz, MeOD) δ 174.43, 168.72, 167.67, 161.60, 153.14,152.75, 140.78, 129.06, 127.74, 125.61, 121.81, 105.37, 70.86, 60.78,57.00, 43.72, 39.35, 33.96, 11.20. MS (ESI) 411.3 (M+H).

VL116 Right Hand Fragment (Representative Method B Synthesis)

2-(trimethylsilyl)ethyl 4-bromobenzylcarbamate

4-Bromobenzylamine hydrochloride (354 mg, 1.59 mmol, 1 eq) was dissolvedin DMF (6.4 mL) and water (2.1 mL) and stirred at room temperature.Triethylamine (0.33 mL, 2.39 mmol, 1.5 eq) and TeocOSu (454 mg, 1.75mmol, 1.1 eq) were then added. After 12 hours, the mixture was dilutedwith EtOAc, washed with 1M HCl, saturated sodium bicarbonate, water andbrine. The organic layer was then dried over sodium sulfate, filtered,and concentrated under reduced pressure. Purification by columnchromatography (10 to 20% EtOAc/hexanes) gave a colorless oil (0.4158 g,1.26 mmol, 79%). 1H NMR (500 MHz, CDCl₃) δ 7.48-7.43 (m, 2H), 7.17 (d,J=8.1 Hz, 2H), 4.94 (s, 1H), 4.31 (d, J=6.0 Hz, 2H), 4.23-4.15 (m, 2H),1.04-0.93 (m, 2H), 0.04 (s, 9H). ¹³C NMR (126 MHz, CDCl₃) δ 156.91,137.95, 131.88, 129.32, 121.43, 63.53, 44.53, 17.92, −1.32. MS (ESI)354.1 (M+H).

2-(trimethylsilyl)ethyl 4-(4-methylthiazol-5-yl)benzylcarbamate

2-(trimethylsilyl)ethyl 4-bromobenzylcarbamate (132 mg, 0.4 mmol, 1 eq),4-methylthiazole-5-carboxylic acid (114.5 mg, 0.8 mmol, 2 eq),tetrabutylammonium chloride hydrate (118 mg, 0.4 mmol, 1 eq), cesiumcarbonate (196 mg, 0.6 mmol, 1.5 eq) and Pd(P(tBu)₃)₂ (40.8 mg, 0.08mmol, 0.2 eq) were dissolved in DMF (4 mL).¹ The reaction was heated to170° C. in a microwave reactor for 16 minutes. The mixture was thencooled to room temperature, diluted with EtOAc and washed thrice withbrine, once with saturated sodium bicarbonate, water, and then brine.The organic layer was dried over sodium sulfate, filtered andconcentrated under reduced pressure. Purification by coulmchromatography (10 to 35% EtOAc/hexanes) gave a colorless oil (61.7 mg,0.177 mmol, 44%). 1H NMR (400 MHz, CDCl₃) δ 8.67 (s, 1H), 7.43-7.37 (m,2H), 7.34 (d, J=8.1 Hz, 2H), 5.09 (s, 1H), 4.39 (d, J=6.0 Hz, 2H),4.28-4.02 (m, 2H), 2.52 (s, 3H), 1.10-0.90 (m, 2H), 0.14-−0.09 (m, 9H).¹³C NMR (101 MHz, CDCl₃) δ 156.98, 150.42, 148.66, 138.76, 131.67,131.18, 129.66, 127.89, 63.46, 44.71, 17.90, 16.18, −1.34. MS (ESI)349.0 (M+H).

(4-(4-methylthiazol-5-yl)phenyl)methanamine

2-(trimethylsilyl)ethyl 4-(4-methylthiazol-5-yl)benzylcarbamate (51.8mg, 0.149 mmol, 1 eq) was dissolvd in acetonittile (6 mL) at roomtemperature. A one molar solution of tetrabutylammonium fluoride in THF(0.45 mL, 0.45 mmol, 3 eq) was added and the solution was stirred for 24hours. The mixture was concentrated under reduced pressure. Purificationby column chromatography (0.5 to 4% 0.5N NH₃ (MeOH)/DCM) gave a lightyellow oil (27.2 mg, 0.133 mmol, 89%). ¹H NMR (500 MHz, MeOD) δ 8.87 (s,1H), 7.44 (s, 4H), 3.85 (s, 2H), 2.47 (s, 3H). ¹³C NMR (126 MHz, MeOD) δ152.77, 149.07, 143.63, 133.42, 131.46, 130.49, 129.05, 46.23, 15.79. MS(ESI) 205.0 (M+H).

Alternate Route:

4-bromobenzonitrile (5.1 g, 28 mmol, 1 eq), 4-methylthiazole (5.56 g, 56mmol, 2 eq) potassium acetate (5.5 g, 56 mmol, 2 eq), palladium (II)acetate (63 mg, 0.28 mmol, 1 mol %) were dissolved in dimethylacetamideand stirred under argon. (CITE JOC, 2009, 74, 1179) The mixture washeated to 150° C. and stirred for 19 hours, then diluted with 500 mLEtOAc, and washed 4 times with 300 mL water. The first wash was thenback extracted with 300 mL EtOAc, and then washed 4 times with 100 mLwater. The combined organic layer was dried over sodium sulfate,filtered and concentrated under vacuum to give a beige solid (5.55 g,27.7 mmol, 99%) that matched the reported spectral data.^([8]) The solidwas then dissolved in MeOH (280 mL) and cooled to 4° C. Cobalt chloride(9.9 g, 41.6 mmol, 1.5 eq) was added, followed by the slow, portionwiseaddition of sodium borohydride (5.2 g, 139 mmol, 5 eq), which wasaccompanied by vigorous bubbling. After 90 minutes, the reaction wasquenched by the addition of water and ammonium hydroxide. The mixturewas extracted 4 times with chloroform, and purified by columnchromatography (10 to 30% 0.5M NH₃ (MeOH)/DCM) to give a darker oil(4.12 g, 20.2 mmol, 73%).

General Solution Phase Synthesis (2S,4R)-allyl4-(tert-butoxy)pyrrolidine-2-carboxylate

(2S,4R)-1-((9H-fluoren-9-yl)methyl) 2-allyl4-(tert-butoxy)pyrrolidine-1,2-dicarboxylate (7.0 g, 15.57 mmol, 1 eq)was dissolved in DCM (156 mL) and cooled to 4° C.Tris(2-aminoethyl)amine (5.8 mL, 38.9 mmol, 2.5 eq) was added, and thesolution was stirred for 1 hour at 4° C. and 4.5 hours at roomtemperature. The mixture was then mixed with silica gel (roughly 20 g),and concentrated under reduced pressure, and purified by columnchromatography (1 to 5% 0.5N NH₃ (MeOH)/DCM) to give an opaque oil (3.44g, 15.1 mmol, 97%). ¹H NMR (400 MHz, MeOH) δ 6.03-5.88 (m, 1H), 5.25(dq, J=17.2, 1.8 Hz, 1H), 5.09 (dq, J=10.5, 1.6 Hz, 1H), 4.33-4.23 (m,1H), 4.06 (dt, J=5.1, 1.6 Hz, 2H), 3.86 (t, J=8.0 Hz, 1H), 3.18 (dd,J=11.4, 5.7 Hz, 1H), 2.70 (dd, J=11.4, 3.8 Hz, 1H), 2.00 (dd, J=8.0, 5.0Hz, 2H), 1.19 (s, 9H). ¹³C NMR (101 MHz, MeOH) δ 175.89, 138.93, 114.88,74.93, 72.93, 63.97, 59.77, 55.44, 40.18, 28.65. MS (ESI) 228.0 (M+H).

(2S,4R)-allyl4-(tert-butoxy)-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylate

(2S,4R)-allyl 4-(tert-butoxy)pyrrolidine-2-carboxylate (0.148 g, 0.65mmol, 1 eq) was dissolved in DMF (6.5 mL) and cooled to 4° C.2-(3-methylisoxazol-5-yl)acetic acid (0.12 g, 0.85 mmol, 1.3 eq), EDC(0.163 g, 0.85 mmol, 1.3 eq), HOBt (0.123 g, 0.91 mmol, 1.4 eq), andDIPEA (0.283 mL, 1.63 mmol, 2.5 eq) were added, and the solution wasallowed to warm slowly to room temperature. After 12 hours, the mixturewas poured into brine and extracted four times with EtOAc. The organiclayer was dried over sodium sulfate, filtered and concentrated underreduced pressure. Purification by column chromatography (1 to 3%MeOH/DCM) gave a light yellow oil (0.2008 g, 0.573 mmol, 88%). ¹H NMR(500 MHz, CDCl₃) δ 6.17 (s, 1H), 5.95-5.85 (m, 1H), 5.29 (ddd, J=13.8,11.7, 1.3 Hz, 2H), 4.69-4.55 (m, 3H), 4.40-4.32 (m, 1H), 3.84-3.75 (m,3H), 3.37 (dd, J=10.0, 4.7 Hz, 1H), 2.27 (s, 3H), 2.15 (ddd, J=18.5,12.0, 5.9 Hz, 2H), 1.18 (s, 9H). ¹³C NMR (126 MHz, CDCl₃) δ 171.87,165.94, 165.63, 160.30, 131.84, 118.72, 104.04, 74.53, 69.55, 65.98,57.96, 54.53, 37.31, 33.58, 28.35, 11.62. MS (ESI) 351.5 (M+H).

(2S,4R)-4-(tert-butoxy)-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid

(2 S,4R)-allyl4-(tert-butoxy)-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylate(1.67 g, 4.77 mmol, 1 eq) was dissolved in THE (48 mL) at roomtemperature. Pd(PPh₃)₄ (0.55 g, 0.48 mmol, 0.1 eq) and morpholine (4.2mL, 48 mmol, 10 eq) were then added. After 35 minutes, the solution wasconcentrated under reduced pressure, redissolved in DCM, and washed fourtimes with 1M HCl (aq). The aqueous layer was then back extracted oncewith DCM. The combined organic layer was then dried over sodium sulfate,filtered and concentrated under reduced pressure. Purification by columnchromatography (1 to 20% MeOH/DCM) gave a yellow solid (1.27 g, 4.1mmol, 86%). ¹H NMR (500 MHz, MeOH) δ 6.23 (s, 1H), 4.47 (t, J=6.0 Hz,2H), 3.94-3.80 (m, 3H), 3.48 (dd, J=10.6, 3.8 Hz, 1H), 2.28-2.11 (m,5H), 1.21 (s, 9H). ¹³C NMR (126 MHz, MeOH) δ 175.53, 168.41, 167.68,161.59, 105.25, 75.57, 71.00, 59.36, 55.81, 38.49, 33.88, 28.48, 11.20.MS (ESI) 311.2 (M+H).

General Method B Representative Procedure (with hydroxyl groupprotection): VL116

(2S,4R)-4-(tert-butoxy)-1-(2-(3-methylisoxazol-5-yl)acetyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

(2S,4R)-4-(tert-butoxy)-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid (53.7 mg, 0.173 mmol, 1.3 eq),(4-(4-methylthiazol-5-yl)phenyl)methanamine (27.2 mg, 0.133 mmol, 1 eq),EDC (33.2 mg, 0.173 mmol, 1.3 eq), and HOBt (23.4 mg, 0.173 mmol, 1.3eq) were dissolved in DMF (3.5 mL) at 4° C. DIPEA (0.07 mL, 0.4 mmol, 3eq) was added, and the solution was allowed to slowly warm to roomtemperature. After 19 hours, the mixture was poured into brine andextracted four times with EtOAc. The organic layer was dried with sodiumsulfate, filtered and concentrated under reduced pressure. Purificationby column chromatography (1 to 5% MeOH/DCM) gave a colorless oil (58.1mg, 0.117 mmol, 88%). ¹H NMR (400 MHz, CDCl₃) δ 8.67 (s, 1H), 7.42-7.27(m, 5H), 6.06 (s, 1H), 4.69 (dd, J=8.4, 2.6 Hz, 1H), 4.59-4.35 (m, 3H),3.82-3.71 (m, 3H), 3.34 (dd, J=9.9, 6.3 Hz, 1H), 2.59-2.46 (m, 4H), 2.25(s, 3H), 1.91 (dd, J=8.2, 4.4 Hz, 1H), 1.25-1.14 (m, 9H). ¹³C NMR (101MHz, CDCl₃) δ 170.70, 167.35, 165.30, 160.24, 150.42, 148.59, 138.09,131.74, 131.05, 129.66, 127.85, 104.19, 74.48, 70.02, 59.12, 54.20,43.25, 35.59, 33.49, 28.38, 16.19, 11.57. MS (ESI) 497.4 (M+H).

(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(VL116)

(2S,4R)-4-(tert-butoxy)-1-(2-(3-methylisoxazol-5-yl)acetyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(58.1 mg, 0.117 mmol) was dissolved in DCM (8 mL). TFA (2 mL, 20%vol/vol) was added and the solution was stirred for 12 hours at roomtemperature, after which it was concentrated under reduced pressure.Purification by column chromatography (1 to 10% 0.5N NH₃ (MeOH)/DCM)gave a colorless oil (28.4 mg, 0.065 mmol, 56%). 1H NMR (400 MHz, MeOH)δ 8.87 (d, J=2.1 Hz, 1H), 7.50-7.34 (m, 4H), 6.23 (s, 1H), 4.57 (t,J=8.0 Hz, 1H), 4.54-4.38 (m, 3H), 3.93 (d, J=2.4 Hz, 2H), 3.81 (dd,J=10.9, 4.3 Hz, 1H), 3.63 (dd, J=7.2, 5.5 Hz, 1H), 2.46 (d, J=8.8 Hz,3H), 2.33-2.20 (m, 4H), 2.10 (ddd, J=13.1, 8.2, 4.7 Hz, 1H). ¹³C NMR(101 MHz, MeOH) δ 174.43, 168.71, 167.66, 161.58, 152.83, 149.04,140.14, 133.39, 131.56, 130.43, 128.88, 105.39, 70.86, 60.78, 57.00,43.65, 39.36, 33.96, 15.81, 11.22. MS (ESI) 441.3 (M+H).

General Method A Representative Procedure: VL133(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid

(2S,4R)-4-(tert-butoxy)-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid (124.9 mg, 0.4 mmol, 1 eq) was dissolved in DCM (18 mL) at roomtemperature. TFA (2 mL, 10%) was added, and the solution was stirred for12 hours. It was then concentrated under reduced pressure and purifiedby column chromatography (4 to 20% MeOH/DCM) to give a yellow oil (99.7mg, 0.39 mmol, 98%). ¹H NMR (500 MHz, MeOD) δ 6.24 (s, 1H), 4.55-4.46(m, 2H), 3.89 (d, J=28.3 Hz, 2H), 3.77 (dd, J=10.9, 4.3 Hz, 1H), 3.62(d, J=11.0 Hz, 1H), 2.36-2.22 (m, 4H), 2.10 (ddd, J=13.1, 8.0, 4.8 Hz,1H). ¹³C NMR (126 MHz, CDCl₃) δ 175.33, 168.51, 167.61, 161.61, 105.28,70.86, 59.33, 56.60, 38.78, 33.85, 11.20. MS (ESI) 255.1 (M+H).

(2S,4R)—N-(4-(1H-pyrrol-3-yl)benzyl)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide(VL133)

(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid (52.6 mg, 0.207 mmol, 1.3 eq),(4-(1H-pyrrol-3-yl)phenyl)methanamine (27.3 mg, 0.159 mmol, 1 eq), EDC(39.7 mg, 0.207 mmol, 1.3 eq) and HOBt (28 mg, 0.207 mmol, 1.3 eq) weredissolved in DMF (4.1 mL) and cooled to 4° C. DIPEA (0.083 mL, 0.477mmol, 3 eq) was added and the solution was allowed to slowly warm toroom temperature. After 16 hours, the mixture was poured into halfsaturated sodium chloride (aqueous) and extracted 3 times with EtOAc.The organic layer was dried over sodium sulfate, filtered andconcentrated under reduced pressure. Purification by columnchromatography (1 to 10% 0.5N NH₃ (MeOH)/DCM) gave an off white solid(41.5 mg, 0.102 mmol, 64%). ¹H NMR (400 MHz, DMSO) δ 8.40 (d, J=6.0 Hz,1H), 7.52-7.39 (m, 2H), 7.22-7.12 (m, 3H), 6.82-6.72 (m, 1H), 6.41 (d,J=1.7 Hz, 1H), 6.24 (s, 1H), 5.17 (d, J=3.9 Hz, 1H), 4.31 (ddd, J=17.1,13.7, 6.4 Hz, 4H), 3.88 (s, 2H), 3.75-3.65 (m, 1H), 3.52-3.41 (m, 1H),2.18 (d, J=18.0 Hz, 3H), 2.12-1.99 (m, 1H), 1.94-1.85 (m, 1H). ¹³C NMR(101 MHz, DMSO) δ 171.36, 166.69, 165.54, 159.38, 135.66, 134.68,127.20, 124.21, 123.00, 118.86, 114.71, 105.22, 103.99, 68.61, 58.76,55.18, 41.63, 38.27, 32.78, 11.00. MS (ESI) 431.5 (M+Na).

For further reference see the following articles and the referencescited therein:

-   (1) Buckley D L et al. J. Am. Chem. Soc 2012, 134, 4465-4468.-   (2) Van Molle I et al. A Chemistry & Biology 2012, 19, 1300-1312-   (3) Buckley, D Angew. Chem. Int. Ed., 2012, 51, 11463-11467-   (4) Buckley, D. L et al. Angew. Chem. 2012, 124, 11630-11634.

Examples Second Set

VL50 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.156 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa white solid (29.8 mg, 0.084 mmol, 54%). 1H NMR (400 MHz, CD₃OD):δ7.34-7.27 (m, 4H); 5.43-5.35 (m, 4H); 3.81-3.78 (dd, J=8 Hz, 4 Hz, 1H);3.61-3.57 (m, 1H); 2.65-2.61 (m, 2H); 2.57-2.51 (m, 2H); 2.28-2.21 (m,1H); 2.08-2.02 (m, 1H). ¹³C NMR (100 MHz, CD₃OD): δ 177.53, 174.74,173.76, 138.75, 133.76, 129.96, 129.49, 70.71, 60.55, 56.47, 43.25,39.33, 30.97, 30.64. MS (ESI) 354.2 (M+H).

VL52 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.156 mmol)according to the Solid Phase Synthesis General Method B. It was isolatedas a white solid (7.7 mg, 0.021 mmol, 14%). 1H NMR (500 MHz, CD₃OD):δ7.41 (d, J=2 Hz, 1H); 7.30 (s, 4H); 6.35-6.34 (dd, J=3 Hz, 2 Hz, 1H);6.26-6.25 (d, J=3 Hz, 1H); 4.49-4.32 (m, 4H); 3.82-3.73 (m, 3H);3.65-2.62 (m, 1H); 2.23-2.22 (m, 1H); 2.09-2.06 (m, 1H).¹³C NMR (125MHz, CD₃OD): δ174.54, 170.58, 149.67, 143.24, 138.68, 133.84, 129.98,129.53, 111.50, 108.98, 70.88, 60.75, 56.95, 43.31, 39.24, 35.58. MS(ESI) 365.2 (M+H), 385.3 (M+Na).

VL73 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.18 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa clear oil (38.9 mg, 0.099 mmol, 55%).¹H NMR (500 MHz, CD₃OD ) δ7.51-6.99 (m, 8H), 4.72 (t, J=8.2, 1H), 4.55-4.33 (m, 3H), 3.60 (dd,J=3.7, 11.3, 1H), 3.19 (dd, J=1.5, 11.3, 1H), 2.36-2.25 (m, 1H),2.21-2.03 (m, 1H).³C NMR (126 MHz, CD₃OD) δ 174.03, 169.66, 138.62,137.31, 133.87, 132.12, 130.92, 130.48, 129.98, 129.56, 129.16, 128.53,70.64, 60.38, 43.39, 39.25, 24.21. MS (ESI) 395.3 (M+H).

VL64 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.156 mmol)according to the Solid Phase Synthesis General Method B. It was isolatedas a white solid (27.5 mg, 0.077 mmol, 49%). ¹H NMR (500 MHz, CD₃OD) δ7.66-7.59 (m, 2H), 7.54-7.22 (m, 7H), 4.75 (t, J=8.6, 1H), 4.55-4.33 (m,3H), 3.85 (dd, J=3.0, 11.5, 1H), 3.43 (d, J=11.5, 1H), 2.38-2.26 (m,1H), 2.14-2.05 (m, 1H).¹³C NMR (126 MHz, CD₃OD) δ 174.72, 172.78,138.73, 137.14, 133.83, 131.74, 129.93, 129.55, 129.49, 128.56, 71.04,60.85, 59.80, 43.34, 39.28. MS (ESI) 359.1 (M+H).

VL69 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.156 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa white solid (26.1 mg, 0.62 mmol, 40%). 1H NMR (500 MHz, DMSO) δ 7.30(dt, J=8.2, 25.1, 4H), 7.20 (dd, J=1.7, 8.3, 1H), 7.13 (d, J=1.7, 1H),7.01 (d, J=8.4, 1H), 4.98 (s, 1H), 4.56 (t, J=8.6, 1H), 4.29 (d, J=2.6,2H), 3.76 (dd, J=15.5, 30.6, 7H), 3.36 (d, J=11.1, 1H), 2.14 (dd, J=7.7,12.8, 1H), 1.96-1.83 (m, 1H). ³C NMR (126 MHz, DMSO) δ 171.91, 168.82,150.39, 148.08, 138.70, 131.10, 128.68, 128.09, 121.00, 111.44, 110.75,99.56, 68.90, 59.33, 59.30, 58.68, 55.57, 41.17, 38.01. MS (ESI) 418.8(M+H).

VL70 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.156 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa colorless oil (31.1 mg, 0.083 mmol, 53%).¹H NMR (500 MHz, CD₃OD) δ7.38-7.17 (m, 6H), 6.85-6.73 (m, 2H), 4.76 (t, J=8.5, 1H), 4.53-4.31 (m,3H), 3.85 (dd, J=3.2, 11.6, 1H), 3.37 (d, J=11.6, 1H), 2.50-2.24 (m,1H), 2.08 (ddd, J=4.3, 9.1, 13.3, 1H).¹³C NMR (126 MHz, CD₃OD) δ 174.72,172.14, 145.18, 138.67, 133.87, 132.18, 129.97, 129.56, 128.80, 122.39,118.87, 117.94, 71.01, 60.29, 58.54, 43.40, 39.40. MS (ESI) 374.5 (M+H).

VL71 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.156 mmol)according to the Solid Phase Synthesis General Method B. It was isolatedas a colorless oil (31.1 mg, 0.080 mmol, 51%). 1H NMR (500 MHz, CD₃OD) δ7.40-7.32 (m, 4H), 7.24 (t, J=7.6, 1H), 7.09 (d, J=7.9, 1H), 7.03 (d,J=7.1, 1H), 4.74 (t, J=8.2, 1H), 4.59-4.33 (m, 3H), 3.54 (d, J=11.0,1H), 3.20 (d, J=11.2, 1H), 2.35 (dd, J=8.7, 12.4, 1H), 2.26 (s, 3H),2.14 (dd, J=4.3, 9.3, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 174.37, 172.44,140.91, 139.29, 138.69, 133.84, 129.93, 129.55, 128.37, 124.30, 121.45,120.64, 70.73, 60.11, 43.36, 39.44, 13.89. MS (ESI) 388.1, 390.3 (M+H).

VL72 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.156 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa yellow oil (31.3 mg, 0.084 mmol, 54%). ¹H NMR (500 MHz, CD₃OD) δ 7.48(d, J=8.3, 2H), 7.30 (s, 4H), 6.79 (d, J=8.3, 2H), 4.79-4.69 (m, 1H),4.53-4.29 (m, 3H), 3.95-3.83 (m, 1H), 3.54 (d, J=11.4, 1H), 2.35-2.24(m, 1H), 2.07 (ddd, J=3.9, 10.1, 13.5, 1H). ¹³C NMR (126 MHz, CD₃OD) δ175.03, 173.02, 149.95, 138.75, 133.80, 130.79, 129.91, 129.54, 126.23,115.89, 71.16, 61.03, 60.12, 43.31, 39.16. MS (ESI) 375.0 (M+H).

VL74 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.18 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa white solid (36.3 mg, 0.092 mmol, 51%). 1H NMR (500 MHz, CD₃OD) δ7.67-7.56 (m, 2H), 7.52-7.44 (m, 2H), 7.34-7.28 (m, 4H), 4.74 (dd,J=7.7, 9.6, 1H), 4.55-4.30 (m, 3H), 3.85 (dd, J=3.5, 11.4, 1H), 3.42 (d,J=11.4, 1H), 2.37-2.28 (m, 1H), 2.15-2.05 (m, 1H). ¹³C NMR (126 MHz,CD₃OD) δ 174.59, 171.54, 138.69, 137.75, 135.66, 133.84, 130.38, 129.92,129.70, 129.55, 71.04, 60.92, 59.75, 43.34, 39.29. MS (ESI) 394.6 (M+H).

VL75 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.18 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa white solid (25.0 mg, 0.066 mmol, 37%). ¹H NMR (500 MHz, CD30OD) δ7.64-6.87 (m, 8H), 4.73 (dd, J=7.7, 9.6, 1H), 4.54-4.31 (m, 3H), 3.84(dd, J=3.5, 11.5, 1H), 3.42 (d, J=11.4, 1H), 2.33 (ddd, J=1.6, 7.6,13.0, 1H), 2.13-2.05 (m, 1H). ³C NMR (126 MHz, CD30OD) δ 174.64, 171.20,163.83 (d, J=246.5), 139.35, 138.72, 133.86, 131.60, 129.94, 129.56,124.51, 118.51 (d, J=21.3), 115.56 (d, J=23.4), 71.02, 60.94, 59.70,43.47, 39.31. MS (ESI) 377.4 (M+H).

VL76 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.18 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa white solid (29.6 mg, 0.067 mmol, 38%). 1H NMR (400 MHz, CD₃OD) δ 7.82(s, 1H), 7.70-7.58 (m, 2H), 7.40 (t, J=7.9, 1H), 7.36-7.18 (m, 4H), 4.73(dd, J=7.9, 9.4, 1H), 4.53-4.31 (m, 3H), 3.82 (dt, J=5.2, 10.4, 1H),3.40 (d, J=11.4, 1H), 2.33 (dd, J=7.6, 13.2, 1H), 2.09 (ddd, J=4.1, 9.7,13.7, 1H). ¹³C NMR (101 MHz, CD₃OD) δ 174.53, 170.95, 139.22, 138.68,134.68, 133.85, 131.56, 131.44, 129.92, 129.56, 127.31, 123.35, 71.02,60.90, 59.70, 43.33, 39.31. MS (ESI) 439.4 (M+H).

VL77 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.18 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa white solid (31.0 mg, 0.081 mmol, 45%). 1H NMR (500 MHz, DMSO) δ 8.07(t, J=1.4, 1H), 8.01-7.95 (m, 1H), 7.93-7.88 (m, 1H), 7.69 (t, J=7.8,1H), 7.40-7.23 (m, 4H), 4.56 (dd, J=8.3, 16.4, 1H), 4.30 (dd, J=8.1,15.4, 3H), 3.79 (dd, J=3.6, 11.0, 1H), 3.24 (d, J=11.0, 1H), 2.23-2.15(m, 1H), 1.92 (ddd, J=4.2, 9.3, 13.2, 1H). ³C NMR (126 MHz, DMSO) δ171.51, 167.29, 138.64, 137.27, 133.88, 132.29, 131.16, 131.08, 129.73,128.68, 128.15, 118.25, 111.40, 68.82, 59.39, 59.36, 58.28, 38.19. MS(ESI) 383.8 (M+H).

VL79 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.18 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa white solid (34.9 mg, 0.090 mmol, 50%). 1H NMR (500 MHz, CD₃OD) δ7.41-7.15 (m, 6H), 7.08-6.90 (m, 2H), 4.73 (dd, J=7.7, 9.6, 1H),4.54-4.31 (m, 3H), 3.87-3.74 (m, 4H), 3.43 (d, J=11.5, 1H), 2.37-2.27(m, 1H), 2.14-2.05 (m, 1H). ³C NMR (126 MHz, CD₃OD) δ 174.72, 172.59,161.07, 138.72, 138.40, 133.83, 130.67, 129.93, 129.56, 120.58, 117.52,113.77, 71.01, 60.82, 59.80, 55.87, 43.33, 39.29. MS (ESI) 389.0 (M+H).

VL80 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.18 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa white solid (41.2 mg, 0.110 mmol, 61%). ¹H NMR (500 MHz, CD₃OD) δ7.36-6.76 (m, 8H), 4.72 (dd, J=7.8, 9.4, 1H), 4.53-4.31 (m, 3H), 3.82(dd, J=3.5, 11.6, 1H), 3.45 (d, J=11.6, 1H), 2.34-2.27 (m, 1H),2.14-2.03 (m, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 174.81, 172.77, 158.73,138.74, 138.36, 133.83, 130.64, 129.93, 129.56, 119.39, 118.62, 115.26,71.01, 60.81, 59.80, 43.46, 39.24. MS (ESI) 375.4 (M+H).

VL81 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.18 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa colorless oil (42.9 mg, 0.091 mmol, 50%). ¹H NMR (500 MHz, CD₃OD) δ7.77-7.27 (m, 6H), 7.04 (d, J=8.3, 1H), 4.71 (t, J=8.2, 1H), 4.56-4.30(m, 3H), 3.59 (dd, J=3.7, 11.2, 1H), 3.17 (d, J=11.3, 1H), 2.37-2.25 (m,1H), 2.19-2.09 (m, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 173.87, 169.41,138.18, 137.15, 133.71, 130.60, 130.36, 130.00, 129.69, 129.56, 129.40,120.48, 70.62, 69.41, 60.48, 43.53, 39.23. MS (ESI) 472.1 (M+H).

VL96 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.155 mmol)according to Solid Phase Synthesis General Method B. It was isolated asa light yellow oil (36.6 mg, 0.102 mmol, 66%). ¹H NMR (500 MHz, CD₃OD) δ8.81 (s, 1H), 8.66 (dd, J=4.6, 1.5 Hz, 2H), 7.60 (dd, J=4.5, 1.6 Hz,2H), 7.32-7.25 (m, 4H), 4.70 (dd, J=9.3, 7.9 Hz, 1H), 4.46 (dd, J=15.3,6.3 Hz, 1H), 4.39 (s, 1H), 4.33 (dd, J=15.4, 5.5 Hz, 1H), 3.78 (dd,J=11.4, 3.5 Hz, 1H), 3.27 (dt, J=3.2, 1.6 Hz, 1H), 2.31 (dd, J=13.2, 7.6Hz, 1H), 2.08 (ddd, J=13.5, 9.6, 4.2 Hz, 1H). ¹³C NMR (126 MHz, CD₃OD) δ174.32, 169.63, 150.65, 145.83, 138.68, 133.88, 129.96, 129.56, 123.32,70.99, 60.88, 59.33, 43.49, 39.33. MS (ESI) 360.5 (M+H).

VL112 was synthesized from Fmoc-Hyp(OWang)-OAllyl resin (0.2 mmol)according to Solid Phase Synthesis General Method A. It was isolated asa cream colored solid (22.6 mg, 0.055 mmol, 28%). 1H NMR (500 MHz,CD₃OD) δ 7.98 (d, J=7.1 Hz, 3H), 7.46 (d, J=8.0 Hz, 2H), 7.28 (s, 1H),6.23 (s, 1H), 4.62-4.39 (m, 4H), 3.93 (d, J=2.9 Hz, 2H), 3.81 (dd,J=10.9, 4.1 Hz, 1H), 3.64 (d, J=11.0 Hz, 1H), 2.33-2.17 (m, 4H),2.15-2.04 (m, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 174.50, 168.72, 167.68,163.41, 161.60, 142.96, 140.75, 129.45, 128.96, 127.53, 127.15, 105.36,70.87, 60.78, 56.99, 43.72, 39.36, 33.95, 11.20. MS (ESI) 410.9 (M+H).

VL115 was synthesized according to General Method B. 1H NMR (500 MHz,CD₃OD) δ 8.87 (s, 1H), 7.46-7.40 (m, 2H), 7.36 (dd, J=8.8, 4.3 Hz, 2H),6.20 (s, 1H), 4.55 (t, J=8.0 Hz, 1H), 4.50 (d, J=6.3 Hz, 1H), 4.48-4.42(m, 2H), 3.92 (d, J=4.5 Hz, 2H), 3.80 (dd, J=10.9, 4.3 Hz, 1H), 3.62 (d,J=11.0 Hz, 1H), 2.48 (d, J=10.2 Hz, 3H), 2.31-2.21 (m, 4H), 2.08 (ddd,J=13.0, 8.2, 4.7 Hz, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 174.48, 168.60,167.70, 161.57, 152.92, 149.26, 140.81, 133.50, 133.09, 130.13, 129.24,129.09, 128.34, 105.35, 70.86, 60.75, 56.95, 43.81, 39.38, 33.92, 15.87,11.23. MS (ESI) 441.4 (M+H).

VL154 was synthesized according to General Method B. 1H NMR (500 MHz,CD₃OD) δ 8.95 (s, 1H), 8.44 (t, J=5.6, 1H), 7.84 (d, J=8.2, 2H), 7.70(d, J=1.9, 1H), 7.35 (d, J=8.2, 2H), 6.19 (s, 1H), 4.56 (t, J=7.9, 1H),4.51 (s, 1H), 4.43 (d, J=5.7, 2H), 3.87 (s, 2H), 3.78 (dd, J=10.9, 4.3,1H), 3.58 (d, J=10.8, 1H), 2.24 (obscured s, 4H), 2.16-2.07 (m, 1H); ¹³CNMR (126 MHz, CD₃OD) δ 176.66, 170.98, 169.94, 164.15, 159.72, 157.62,142.33, 136.89, 131.58, 130.33, 117.00, 108.05, 73.36, 63.27, 59.60,46.75, 41.78, 36.80, 14.39; TLC: (EtOAC) R_(f)=0.5; LRMS (ESI) 427.6(M+H)⁺.

VL155 was synthesized according to General Method B. ¹H NMR (500 MHz,CD₃OD) δ 8.87 (d, J=5.2, 1H), 8.54 (t, J=5.7, 1H), 8.07 (s, 1H), 7.56(d, J=8.2, 2H), 7.36 (d, J=8.2, 2H), 6.20 (s, 1H), 4.56 (t, J=8.0, 1H),4.51 (s, 1H), 4.42 (qd, J=5.5, 15.5, 2H), 3.78 (dt, J=9.2, 18.5, 1H),3.60 (d, J=11.1, 1H), 2.28-2.21 (m, 4H), 2.10 (ddd, J=4.7, 8.0, 13.0,1H); ¹³C NMR (126 MHz, CD₃OD) δ 176.72, 171.03, 169.98, 164.12, 142.98,142.96, 133.45, 132.34 131.86, 130.07, 108.02, 100.0, 73.37, 63.29,59.60, 46.52, 41.81, 36.74, 14.27. TLC: (EtOAC) R_(f)=0.5; LRMS (ESI)427.4 (M+H)⁺.

VL118 was synthesized according to General Method A. ¹H NMR (500 MHz,CD₃OD) δ 7.37-7.31 (m, 2H), 7.27 (d, J=7.7 Hz, 1H), 7.22 (d, J=7.5 Hz,1H), 6.74-6.68 (m, 1H), 6.20 (s, 1H), 6.14 (dd, J=3.5, 1.8 Hz, 1H),6.10-6.05 (m, 1H), 4.55 (t, J=8.0 Hz, 1H), 4.49 (s, 1H), 4.45-4.39 (m,2H), 3.89 (t, J=8.4 Hz, 1H), 3.79 (dd, J=10.9, 4.3 Hz, 1H), 3.67-3.55(m, 5H), 2.26-2.22 (m, 4H), 2.07 (ddd, J=13.0, 8.1, 4.7 Hz, 1H). ¹³C NMR(126 MHz, CD₃OD) δ 174.35, 168.57, 167.66, 161.57, 139.94, 135.44,135.24, 129.58, 128.35, 128.22, 126.62, 124.93, 109.56, 108.54, 105.37,70.84, 60.72, 56.91, 44.04, 39.36, 35.34, 33.88, 11.23. MS (ESI) 422.8(M+H).

VL119 was synthesized according to General Method A. ¹H NMR (500 MHz,CD₃OD) δ 7.38-7.30 (m, 4H), 6.76-6.67 (m, 1H), 6.23 (s, 1H), 6.10 (dd,J=3.5, 1.8 Hz, 1H), 6.09-6.05 (m, 1H), 4.56 (t, J=8.1 Hz, 1H), 4.51 (s,1H), 4.47-4.39 (m, 2H), 3.93 (d, J=3.0 Hz, 2H), 3.81 (dd, J=10.9, 4.3Hz, 1H), 3.63 (q, J=5.8 Hz, 4H), 2.31-2.22 (m, 4H), 2.10 (ddd, J=13.0,8.1, 4.7 Hz, 1H). ¹³C NMR (101 MHz, ˜1:1 CD₃OD:CDCl₃) δ 172.63, 167.39,166.19, 160.68, 136.80, 132.70, 129.06, 128.99, 127.75, 124.09, 108.75,107.93, 104.62, 69.88, 59.64, 56.15, 43.36, 37.98, 35.19, 33.53, 11.37.MS (ESI) 423.6 (M+H).

VL131 was synthesized according to General Method B. 1H NMR (400 MHz,CD₃OD) δ 9.02 (d, J=5.2 Hz, 1H), 8.02 (d, J=8.4 Hz, 2H), 7.41 (d, J=8.4Hz, 2H), 6.17 (s, 1H), 4.52-4.38 (m, 4H), 3.84 (s, 2H), 3.76 (dd,J=10.8, 4.3 Hz, 1H), 3.56 (d, J=9.5 Hz, 1H), 2.30-2.18 (m, 4H), 2.14(td, J=8.1, 3.9 Hz, 1H). ¹³C NMR (101 MHz, CD₃OD) δ 173.25, 167.86,167.57, 166.44, 166.18, 160.85, 142.53, 128.33, 128.14, 125.56, 104.77,70.04, 59.87, 56.31, 43.52, 38.30, 33.60, 11.39. MS (ESI) 413.3 (M+H).

VL138 was synthesized according to General Method B. 1H NMR (400 MHz,CD₃OD) δ 7.42 (d, J=8.2 Hz, 2H), 7.32-7.24 (m, 2H), 6.24 (s, 1H),4.69-4.33 (m, 5H), 3.94 (d, J=3.0 Hz, 1H), 3.82 (dd, J=10.9, 4.3 Hz,1H), 3.64 (d, J=11.1 Hz, 1H), 2.38 (s, 3H), 2.31-2.24 (m, 4H), 2.23 (s,3H), 2.10 (ddd, J=13.1, 8.2, 4.7 Hz, 1H). ³C NMR (126 MHz, CD₃OD) δ174.41, 168.72, 167.67, 166.87, 161.59, 160.02, 139.46, 130.38, 129.37,128.89, 117.72, 105.38, 70.87, 60.78, 57.01, 43.74, 39.37, 33.97, 11.38,11.20, 10.66. MS (ESI) 438.6 (M+H).

VL139 was synthesized according to General Method B. 1H NMR (400 MHz,˜1:1 CD₃OD:CDCl₃) δ 7.89 (s, 2H), 7.49 (d, J=8.1 Hz, 2H), 7.28 (d, J=7.9Hz, 2H), 6.20 (s, 1H), 4.54 (dd, J=17.4, 9.5 Hz, 2H), 4.39 (d, J=5.3 Hz,2H), 3.93-3.46 (m, 4H), 2.32-2.16 (m, 4H), 2.16-2.05 (m, 1H). ³C NMR(126 MHz, ˜1:1 CD₃OD:CDCl₃) δ 173.80, 168.23, 167.21, 161.29, 137.35,132.66, 129.24, 128.77, 126.60, 126.48, 105.16, 70.53, 60.42, 56.73,43.71, 39.00, 33.85, 11.30. MS (ESI) 410.0 (M+H).

VL152 was synthesized according to General Method B. 1H NMR (400 MHz,CD₃OD) δ 7.38 (d, J=8.2 Hz, 2H), 7.25 (d, J=8.2 Hz, 2H), 6.17 (d, J=55.2Hz, 1H), 4.65-4.30 (m, 4H), 4.05-3.72 (m, 3H), 3.64 (d, J=11.1 Hz, 1H),2.32-2.19 (m, 10H), 2.10 (ddd, J=13.1, 8.2, 4.7 Hz, 1H). ¹³C NMR (101MHz, CD₃OD) δ 174.37, 168.72, 167.67, 161.59, 143.18, 138.27, 132.85,130.54, 129.12, 128.64, 105.40, 70.86, 60.78, 57.01, 43.80, 39.38,33.96, 11.21, 11.07. MS (ESI) 438.5 (M+H).

VL158 was synthesized according to General Method B. 1H NMR (500 MHz,CD₃OD) δ 8.03 (s, 1H), 7.62 (t, J=8.7 Hz, 2H), 7.43 (d, J=6.6 Hz, 1H),7.33 (t, J=6.5 Hz, 2H), 6.19 (s, 1H), 4.62-4.48 (m, 2H), 4.48-4.32 (m,2H), 3.93-3.68 (m, 3H), 3.58 (s, 1H), 2.29-2.19 (m, 4H), 2.11 (ddd,J=13.0, 8.0, 4.8 Hz, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 173.59, 168.01,166.92, 161.11, 138.84, 135.95, 130.48, 129.06, 128.64, 125.89, 116.23,105.04, 70.35, 60.24, 56.60, 43.56, 38.76, 33.78, 11.33. MS (ESI) 410.1(M+H).

VL160 was synthesized according to General Method B. ¹H NMR (500 MHz,CD₃OD) δ 7.68 (d, J=8.1 Hz, 2H), 7.63 (s, 1H), 7.33 (d, J=8.0 Hz, 2H),6.63 (s, 1H), 6.19 (s, 1H), 4.54 (t, J=8.0 Hz, 1H), 4.47 (s, 1H), 4.38(d, J=4.6 Hz, 2H), 3.89 (d, J=3.1 Hz, 2H), 3.78 (dd, J=10.9, 4.3 Hz,1H), 3.60 (d, J=11.1 Hz, 1H), 2.28-2.14 (m, 4H), 2.06 (ddd, J=13.0, 8.1,4.7 Hz, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 174.32, 168.69, 167.67, 161.61,139.59, 132.31, 129.33, 128.86, 127.00, 126.87, 105.49, 105.38, 70.84,60.78, 56.97, 43.80, 39.35, 33.95, 11.19. MS (ESI) 409.2 (M+H), 431.8(M+Na).

(2S,4R)-(9H-fluoren-9-yl)methyl4-(tert-butoxy)-2-((4-chlorobenzyl)carbamoyl)pyrrolidine-1-carboxylate

Fmoc-Hyp(OtBu)-OH (1.23 g, 3 mmol, 1 eq) was dissolved in DCM (15 mL)and cooled to 4° C. EDC (0.69 g, 3.6 mmol, 1.2 eq) and HOBt (0.49 g, 3.6mmol, 1.2 eq) were then added. After 20 minutes, 4-chlorobenzylamine(0.48 mL, 3.9 mmol, 1.3 eq) was added and the solution was allowed towarm slowly to room temperature. After 15 hours, the mixture was dilutedwith EtOAc and washed with 1M HCl, sodium bicarbonate, water and brine.The organic layer was dried with sodium sulfate, filtered and condensed.Purification by column chromatography (25 to 100% EtOAc/hexanes) gave awhite foam (1.42 g, 2.66 mmol, 89%). ¹H NMR (400 MHz, CDCl₃) δ 7.77 (d,J=7.2 Hz, 2H), 7.57 (s, 2H), 7.40 (t, J=7.3 Hz, 2H), 7.31 (t, J=7.3 Hz,2H), 7.17 (dd, J=27.2, 19.5 Hz, 4H), 4.58-3.94 (m, 7H), 3.60 (d, J=6.7Hz, 1H), 3.31 (d, J=6.6 Hz, 1H), 2.50 (s, 1H), 1.96 (s, 1H), 1.28-1.10(m, 9H). ¹³C NMR (101 MHz, CDCl₃) δ 171.54, 156.13, 143.74, 141.32,136.81, 133.02, 128.79, 128.71, 127.83, 127.12, 125.04, 120.07, 74.15,69.63, 67.87, 59.17, 53.26, 47.10, 42.72, 36.34, 28.31. MS (ESI) 534.8(M+H).

(2S,4R)-4-(tert-butoxy)-N-(4-chlorobenzyl)pyrrolidine-2-carboxamide

(2S,4R)-(9H-fluoren-9-yl)methyl4-(tert-butoxy)-2-((4-chlorobenzyl)carbamoyl)pyrrolidine-1-carboxylate(0.5 g, 0.94 mmol, 1 eq) was dissolved in DCM (15 mL) and cooled to 4°C. Tris(2-aminoethyl)amine (0.35 mL, 2.34 mmol, 2.5 eq) was addedslowly, dropwise. After 30 minutes, the reaction was warmed to roomtemperature and stirred for an additional 14 hours. It was loadeddirectly onto a silica column, and purified by column chromatography (1to 7% 0.5N methanolic ammonia/DCM) to give a white solid (0.2871 g, 0.92mmol, 98%). ¹H NMR (400 MHz, CD30OD) δ 7.27 (dd, J=20.1, 8.4 Hz, 4H),4.35 (s, 2H), 4.22 (s, 1H), 3.84 (t, J=8.0 Hz, 1H), 3.08 (dd, J=11.4,5.1 Hz, 1H), 2.76 (dd, J=11.4, 2.8 Hz, 1H), 2.14-1.98 (m, 1H), 1.97-1.81(m, 1H), 1.17 (s, 9H). ¹³C NMR (126 MHz, CD30OD) δ 176.48, 138.81,133.83, 130.00, 129.52, 74.76, 73.37, 60.80, 55.61, 43.04, 40.76, 28.67.

General Method C: Representative Procedure: VL156

1H-Imidazol-1-ylacetic acid (20.6 mg, 0.163 mmol, 1.3 eq), EDC (31.2 mg,0.163 mmol, 1.3 eq) and HOBt (22 mg, 0.163 mmol, 1.3 eq) were dissolvedin DCM (2.5 mL) and DMF (0.4 mL) at room temperature in a 1 dram vial.After stirring for 15 minutes, DIPEA (0.055 mL, 0.313 mmol, 2.5 eq) wasadded, followed by(2S,4R)-4-(tert-butoxy)-N-(4-chlorobenzyl)pyrrolidine-2-carboxamide(38.9 mg, 0.125 mmol, 1 eq) after an additional 30 minutes. The mixturewas stirred for 14 hours, then diluted with EtOAc and washed with brine.The organic layer was dried over sodium sulfate, filtered and condensed.Purification by column chromatography (1 to 10% MeOH/DCM) gave a whitesolid, which was used directly in the following step. ¹H NMR (400 MHz,CD₃OD) δ 7.65 (s, 1H), 7.28 (td, J=10.9, 8.4 Hz, 4H), 7.06 (d, J=43.6Hz, 2H), 4.99 (dd, J=38.1, 17.1 Hz, 2H), 4.51 (t, J=6.6 Hz, 2H), 4.35(q, J=15.4 Hz, 2H), 3.86 (dd, J=10.2, 5.6 Hz, 1H), 3.45 (dd, J=10.3, 4.1Hz, 1H), 2.22-2.02 (m, 2H), 1.21 (d, J=13.8 Hz, 9H). MS (ESI) 419.7(M+H).

The white solid was dissolved in DCM (9 mL) at room temperature. TFA (1mL) was added and the mixture was stirred for 12 hours and condensed.Purification by column chromatography (1 to 20% 0.5 N methanolicammonia/DCM) gave a white solid (39.8 mg, 0.11 mmol, 88% over 2 steps.¹H NMR (400 MHz, CD₃OD) δ 8.73 (s, 1H), 7.47 (d, J=16.9 Hz, 2H), 7.26(s, 4H), 5.25 (dd, J=37.5, 16.9 Hz, 2H), 4.56 (t, J=7.9 Hz, 2H),4.44-4.27 (m, 2H), 3.82 (dd, J=10.8, 4.1 Hz, 1H), 3.63 (d, J=10.8 Hz,1H), 2.36-2.22 (m, 1H), 2.07 (ddd, J=13.1, 8.3, 4.6 Hz, 1H). ¹³C NMR(126 MHz, CD₃OD) δ 174.14, 166.34, 138.56, 138.20, 133.87, 129.97,129.49, 124.55, 121.47, 70.94, 61.00, 55.75, 51.33, 43.35, 39.21. MS(ESI) 364.8 (M+H).

VL120 was synthesized according to General Method C. ¹H NMR (500 MHz,CD₃OD) δ 8.67 (d, J=1.2 Hz, 1H), 7.36 (s, 1H), 7.29 (d, J=9.2 Hz, 4H),4.62-4.47 (m, 2H), 4.47-4.24 (m, 2H), 3.87 (ddd, J=18.3, 15.1, 10.8 Hz,3H), 3.66 (d, J=11.0 Hz, 1H), 2.37-2.20 (m, 1H), 2.07 (ddd, J=13.1, 8.4,4.6 Hz, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 174.56, 169.45, 138.62, 135.15,133.89, 129.98, 129.72, 129.52, 118.80, 70.89, 60.70, 56.84, 43.35,39.46, 31.70. MS (ESI) 362.3 (M+H).

VL157 was synthesized according to General Method C. 1H NMR (500 MHz,CD₃OD) δ 7.49 (s, 1H), 7.39 (s, 1H), 7.34-7.22 (m, 4H), 4.52 (t, J=8.1Hz, 1H), 4.50-4.45 (m, 1H), 4.37 (dt, J=22.8, 15.4 Hz, 2H), 3.87-3.80(m, 3H), 3.77 (dd, J=11.0, 4.2 Hz, 1H), 3.66-3.52 (m, 3H), 2.30-2.18 (m,1H), 2.04 (ddd, J=13.1, 8.3, 4.7 Hz, 1H). ¹³C NMR (126 MHz, CD₃OD) δ174.67, 172.70, 140.18, 138.71, 133.81, 131.62, 129.95, 129.51, 115.17,70.91, 60.65, 56.93, 43.29, 39.28, 38.76, 31.51. MS (ESI) 377.0 (M+H).

VL173 was synthesized according to General Method C. ¹H NMR (400 MHz,CDCl₃/CD₃OD) δ 7.63-7.53 (m, 3H); 7.49 (d, J=7.6 Hz, 1H); 7.26 (q, J=8.3Hz, 4H); 4.60-4.51 (m, 2H); 4.42-4.32 (m, 2H); 3.84-3.75 (m, 3H); 3.59(d, J=11.3 Hz, 1H); 3.42-3.32 (m, 1H); 2.29-2.19 (m, 1H); 2.17-2.08 (m,1H). ¹³C NMR (100 MHz, CDCl₃/CD₃OD) δ 173.4, 170.7, 137.4, 136.8, 134.8,133.6, 131.1, 129.9, 129.3, 129.0, 119.2, 112.7, 70.1, 59.8, 56.3, 43.0,41.1, 38.4. TLC (10% MeOH in CH₂Cl₂), R_(f) 0.38 (UV, CAM), MS (ESI+):calculated for C₂₁H₂₁N₃O₃Cl [M+H]⁺ 398.1, found 398.2.

Azidoacetic Acid

To a solution of ethyl azidoacetate (530 mg, 4.107 mmol) in THF-H₂O (12mL/12 mL) at rt was added LiOH.H₂O (345 mg, 8.214 mmol). The reactionmixture was stirred at rt for 17 h, evaporated, and diluted with H₂O (10mL), cooled to 0° C., and adjusted to pH 4 with 1N—HCl.

The resulting mixture was extracted twice with diethyl ether, washedwith brine, dried over anhydrous Na₂SO₄, and evaporated. The concentratewas purified by short column chromatography (eluting with 100% hexaneinitially, grading to 2% ethyl acetate in hexane) on silica gel to giveazidoacetic acid 1 (372 mg, 89%) as a pale-yellow oil. ¹H NMR (500 MHz,CDCl₃) δ 9.73 (brs, 1H), 3.97 (s, 2H). ¹³C NMR (125 MHz, CD₃OD) δ 174.2,50.0.

(2S,4R)-1-(2-azidoacetyl)-4-(tert-butoxy)-N-(4-chlorobenzyl)pyrrolidine-2-carboxamide

To a solution of azidoacetic acid (32 mg, 0.315 mmol) in CH₂Cl₂-DMF (1.5mL/1.5 mL)) at room temperature were added(2S,4R)-4-(tert-butoxy)-N-(4-chlorobenzyl)pyrrolidine-2-carboxamide (93mg, 0.300 mmol), DIPEA (0.19 mL, 1.080 mmol), and HOBt (48 mg, 0.360mmol). The mixture was cooled to 0° C., and then EDC (69 mg, 0.360 mmol)was added to the mixture at 0° C. The reaction mixture was allowed towarm to rt, stirred at rt for 17 h, and cooled to 0° C. The resultingmixture was quenched with H₂O (5 mL) and extracted twice withethylacetate. The combined extracts were washed with brine, dried overanhydrous Na₂SO₄, filtered, and evaporated. The concentrate was purifiedby column chromatography (eluting with 5% ethyl acetate in hexaneinitially, grading to 40% ethyl acetate in hexane) on silica gel toafford the coupled product (110 mg, 93%). 1H NMR (400 MHz, CDCl₃) δ 7.31(brs, 1H), 7.27 (d, J=8.5 Hz, 2H), 7.18 (d, J=8.5 Hz, 2H), 4.66 (dd,J=8.4, 2.2 Hz, 1H), 4.58-4.52 (m, 1H), 4.41 (dd, J=15.1, 6.1 Hz, 1H),4.30 (dd, J=15.1, 5.8 Hz, 1H), 3.87 (dd, J=16.0, 16.0 Hz, 1H), 3.84 (dd,J=16.0, 16.0 Hz, 1H), 3.61 (dd, J=9.8, 7.0 Hz, 1H), 3.14 (dd, J=9.8, 6.4Hz, 1H), 2.50 (ddd, J=12.6, 6.3, 2.3 Hz, 1H), 1.86 (dt, J=12.6, 8.2 Hz,1H), 1.19 (s, 9H). ¹³C NMR, 100 MHz, CDCl₃) δ 170.3, 167.4, 162.5,136.5, 133.1, 128.8, 128.7, 74.3, 69.9, 59.0, 52.7, 50.9, 42.8, 36.4,35.1, 31.4, 28.2. MS (ESI) [M+H]⁺ 394.3.

(2S,4R)-4-(tert-butoxy)-N-(4-chlorobenzyl)-1-(2-(4-(methoxymethyl)-1H-1,2,3-triazol-1-yl)acetyl)pyrrolidine-2-carboxamide

To a solution of methyl propargyl ether (7 mg, 0.067 mmol) and(2S,4R)-1-(2-azidoacetyl)-4-(tert-butoxy)-N-(4-chlorobenzyl)pyrrolidine-2-carboxamide(25 mg, 0.0636 mmol) in t-BuOH-H₂O (1:1, 1 mL) and THF (1 mL) at rt wereadded CuSO₄.5H₂O (1.5 mg, 0.006 mmol) and sodium ascorbate (1.0 M inH₂O, 2 drops). The reaction mixture was stirred at rt for 19 h andevaporated. The residue was diluted with H₂O (5 mL) and the mixture wasextracted three times with ethyl acetate. The combined extracts werewashed with brine, dried over Na₂SO₄, filtered, and concentrated. Thecrude residue was purified by flash chromatography on silica gel to givethe desired triazole (25 mg, 85%). ¹H NMR (500 MHz, CD₃OD) δ 7.91 (s,1H), 7.30-7.22 (m, 4H), 5.43 (d, J=16.8 Hz, 1H), 5.34 (d, J=16.7 Hz,1H), 4.54 (s, 2H), 4.53-4.49 (m, 2H), 4.37 (d, J=15.4 Hz, 1H), 4.31 (d,J=15.4 Hz, 1H), 3.90 (dd, J=10.3, 5.6 Hz, 1H), 3.49 (dd, J=10.4, 4.3 Hz,1H), 3.37 (s, 3H), 2.19-2.13 (m, 1H), 2.11-2.07 (m, 1H), 1.22 (s, 9H).¹³C NMR (asterisk denotes the signals of the minor rotamer, 125 MHz,CD₃OD) δ 174.1, *173.4, *166.9, 166.7, 145.7, *138.6, 138.5, *134.2,133.8, *130.4, 129.9, *129.7, 129.5, 126.7, 75.6, *75.5, 71.1, *69.2,66.3, *66.2, 60.8, *60.3, 58.4, *58.3, *55.5, 54.8, 52.6, *51.9, *43.7,43.3, *41.1, 38.7, 28.5. MS (ESI) [M+H]⁺ 464.2.

To a stirred solution of the corresponding t-butyl ether (22 mg, 0.0475mmol) in CH₂Cl₂ (1.5 mL) at 0° C. was added TFA (0.2 mL). The reactionmixture was stirred at rt for 12 h and concentrated. The residue waschromatographed (eluting with 100% CH₂Cl₂ initially, grading to 7% CH₃OHin CH₂Cl₂) on silica gel to provide 5 (18.5 mg, 96%). ¹H NMR (500 MHz,CD₃OD/CDCl₃=2:1) d 8.83 & 8.46 (due to the rotamers, both t, J=5.7 Hz,1H), 7.85 & 7.76 (due to the rotamers, both s, 1H), 7.23 (d, J=8.5 Hz,2H), 7.18 (d, J=8.5 Hz, 2H), 5.36 (d, J=16.6 Hz, 1H), 5.27 (d, J=16.6Hz, 1H), 4.54 (s, 2H), 4.54-4.50 (m, 2H), 4.33 & 4.32 (due to therotamers, both s, 2H), 3.77 (dd, J=10.8, 4.2 Hz, 1H), 3.60 (d, J=10.8Hz, 1H), 3.37 & 3.36 (due to the rotamers, both s, 3H), 2.26-2.21 (m,1H), 2.09-2.04 (m, 1H). ¹³C NMR (asterisk denotes the signals of theminor rotamer, 125 MHz, CD₃OD/CDCl₃=2:1) d *173.4, 173.3, *166.2, 165.9,145.2, 137.5, *133.9, 133.5, 129.9, 129.4, 129.1, 126.0, *125.9, 70.4,*68.5, 66.0, *65.9, *60.4, 60.3, 58.4, *58.3, *56.1, 55.4, 52.2, *51.5,*43.2, 43.1, *41.1, 38.4. MS (ESI) [M+H]⁺ 408.3. TLC (10% MeOH inCH₂Cl₂), R_(f) 0.48 (UV, CAM).

VL167 was synthesized according to General Method C. 1H NMR (500 MHz,CDCl₃) δ 7.86 (s, 1H), 7.71 (d, J=9.5 Hz, 1H), 7.61 (d, J=7.4 Hz, 2H),7.56 (d, J=7.6 Hz, 1H), 7.51 (d, J=7.6 Hz, 1H), 7.42 (dd, J=7.4, 7.4 Hz,2H), 7.38-7.30 (m, 2H), 7.30-7.26 (m, 3H), 4.66-4.42 (m, 2H), 4.42-4.27(m, 2H), 3.85 (dd, J=11.4, 3.5 Hz, 1H), 3.49 (d, J=11.4 Hz, 1H),2.33-2.29 (m, 1H), 2.15-2.09 (m, 1H). ¹³C NMR (asterisk denotes thesignals of the minor rotamer, 125 MHz, CDCl₃) δ 172.5, 170.6, 162.7,140.7, 139.3, 136.4, 135.6, 131.9, 128.2, *128.1, 128.0, 127.9, 127.6,*127.5, 126.8, 126.1, *126.0, 125.2, 125.1, 68.9, *67.4, *60.2, 58.7,57.8, 41.5, *39.5, 37.2, 35.2, 30.0. MS (ESI) [M+H]⁺ 435.5.

VL216 was synthesized according to General Method C.

(2S,4R)-(9H-fluoren-9-yl)methyl4-(tert-butoxy)-2-((4-(oxazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate

To a round bottom flask with stir bar was charged(2S,4R)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-(tert-butoxy)pyrrolidine-2-carboxylicacid (0.587, 1.4 mmol 1.0 equiv)′ EDC (380 mg, 2.0 mmol, 1.4 equiv),HOBt (310 mg, 2.0 mmol, 1.4 equiv) and(4-(oxazol-5-yl)phenyl)methanamine (250 mg, 1.4 mmol, 1.0 equiv). Uponstirring for 18 h the reaction was diluted with 25 ml DCM, and washedwith citric acid (2×50 mL), and saturated NaHCO₃ (2×50 mL). The organiclayer was dried with Na₂SO₄, concentrated down then purified via silicagel chromatography (DCM to 2% MeOH (0.5 N NH₃) to yield 515 mg (65%yield) of product as a viscous oil 1H NMR (400 MHz, CDCl₃) δ 7.89 (s,1H), 7.85-7.70 (m, 2H), 7.68-7.49 (m, 4H), 7.47-7.37 (m, 2H), 7.35-7.20(m, 5H), 4.54-4.35 (m, 4H), 4.35-4.14 (m, 2H), 3.72-3.58 (m, 1H),3.49-3.27 (m, 1H), 2.53 (s, 1H), 2.00 (dd, J=8.1, 20.2, 1H), 1.25 (s,9H); TLC: (9:1 DCM:MeOH (0.5 N NH₃)) R_(f)=0.5; LRMS (ESI) 565.9 (M+H)⁺.

(2S,4R)-1-(3-ethoxybenzoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

To (2S,4R)-(9H-fluoren-9-yl)methyl4-(tert-butoxy)-2-((4-(oxazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate (2.5 g, 3.61 mmol, 1.0 equiv) in 36 mL DCM wascharged tris(2-aminoethyl)amine mol, 9.0 mmol, 2.5 equiv). □(400 uponstirring for 3 h the cloudy mixture was diluted with silica gel andconcentrated down. The material was then dry loaded to a silica gelcolumn and purified (DCM to 5% MeOH (0.5 N NH₃) in DCM) to yield 820 mg(67% yield) as a white solid. ¹H NMR (501 MHz, CDCl₃) δ 8.02 (s, 1H),7.90 (s, 1H), 7.60 (d, J=8.1, 2H), 7.33-7.29 (m, 3H), 4.44 (d, J=6.1,2H), 4.21-4.07 (m, 1H), 3.97 (dd, J=7.2, 8.7, 1H), 2.87 (d, J=11.6, 1H),2.80 (dd, J=4.3, 11.6, 1H), 2.17 (dd, J=10.2, 12.4, 1H), 2.11-1.94 (m,1H), 1.16 (s, 9H); ¹³C NMR (126 MHz, CDCl₃) δ 174.88, 151.27, 150.40,139.33, 128.07, 126.79, 124.63, 121.43, 73.65, 72.30, 59.95, 54.97,42.51, 39.27, 28.38; TLC: (9:1 DCM:MeOH (0.5 N NH₃)) R_(f)=0.42; LRMS(ESI) 344.2 (M+H)⁺.

¹H NMR (400 MHz, CD₃OD) δ 7.42-7.28 (m, 4H), 7.28-7.12 (m, 2H),7.10-6.77 (m, 2H), 4.71 (dt, J=30.7, 15.3 Hz, 1H), 4.58-4.30 (m, 3H),4.18-3.92 (m, 2H), 3.87-3.77 (m, 1H), 3.44 (d, J=11.4 Hz, 1H), 2.38-2.24(m, 1H), 2.15-2.03 (m, 1H), 1.55-1.23 (m, 3H). ¹³C NMR (126 MHz, CD₃OD)δ 174.72, 172.63, 160.34, 138.72, 138.35, 133.82, 130.65, 130.11,129.92, 129.55, 120.47, 118.08, 114.27, 113.89, 71.01, 64.71, 60.81,59.80, 43.33, 39.29, 15.09. MS (ESI) 403.2 (M+H).

General Method D: Representative Procedure: VL217

(2S,4R)-4-(tert-butoxy)-1-(3-ethoxybenzoyl)-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide

3-Ethoxybenzoic acid (13.3 mg, 0.08 mmol, 1 eq), EDC (16.9 mg, 0.088mmol, 1.1 eq) and HOBt (11.9 mg, 0.88 mmol, 1.1 eq) were dissolved inDCM (0.8 mL) at room temperature. DIPEA (0.0279 mL, 0.16 mmol, 2 eq) wasadded, followed by(2S,4R)-4-(tert-butoxy)-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide(33.0 mg, 0.096 mmol, 1.2 eq). The solution was stirred for 21 hoursthen diluted with EtOAc and washed with 10% citric acid, saturatedsodium bicarbonate and brine. The organic layer was dried over sodiumsulfate, filtered and condensed. Purification by column chromatography(1 to 5% MeOH/DCM) gave a colorless oil (36.1 mg, 0.073 mmol, 92%). ¹HNMR (400 MHz, CDCl₃) δ 7.90 (s, 1H), 7.61 (dd, J=16.6, 6.9 Hz, 3H),7.38-7.27 (m, 4H), 6.98 (dd, J=16.0, 6.4 Hz, 3H), 4.92 (dd, J=8.3, 4.7Hz, 1H), 4.48 (d, J=6.0 Hz, 2H), 4.43-4.31 (m, 1H), 4.03 (q, J=7.0 Hz,2H), 3.61 (dd, J=10.9, 5.7 Hz, 1H), 3.31 (dd, J=10.9, 4.4 Hz, 1H),2.73-2.55 (m, 1H), 2.05-1.92 (m, 1H), 1.40 (t, J=7.0 Hz, 3H), 1.13 (s,9H). MS (ESI) 492.4 (M+H).

(2S,4R)-4-(tert-butoxy)-1-(3-ethoxybenzoyl)-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide(36.1 mg, 0.073 mmol, 1 eq) was dissolved in DCM (9 mL) at roomtemperature. TFA (1 mL) was added and the solution was stirred for 13hours, then condensed. Purification by column chromatography (1 to 10%MeOH/DCM) gave a colorless oil (22.9 mg, 0.053 mmol, 72%). 1H NMR (400MHz, CD₃OD) δ 8.24 (d, J=12.0 Hz, 1H), 7.65 (dd, J=28.0, 8.3 Hz, 2H),7.47 (dd, J=18.8, 10.6 Hz, 3H), 7.23 (ddd, J=9.4, 4.6, 4.1 Hz, 3H),7.09-6.87 (m, 2H), 4.75 (dd, J=9.6, 7.7 Hz, 1H), 4.48 (dd, J=49.7, 15.5Hz, 3H), 4.06 (q, J=7.0 Hz, 2H), 3.84 (dd, J=11.5, 3.5 Hz, 1H), 3.44 (d,J=11.5 Hz, 1H), 2.42-2.29 (m, 1H), 2.21-2.05 (m, 1H), 1.36 (dt, J=24.0,7.0 Hz, 3H). ¹³C NMR (101 MHz, CD₃OD) δ 174.78, 172.66, 160.35, 153.14,152.74, 140.85, 138.38, 130.66, 129.00, 127.71, 125.62, 121.77, 120.50,118.08, 114.30, 71.02, 64.71, 60.85, 59.82, 43.72, 39.32,

(2S,4R)-tert-butyl4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate

(2S,4R)-1-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid(366 mg, 1.58 mmol, 1 equiv.) was dissolved in 15 mL DMF and chargedwith EDC (380 mg, 2.0 mmol 1.3 equiv.), and HOBt (310 mg, 2.0 mmol, 1.5equiv) after 5 minutes of stirring(4-(4-methylthiazol-5-yl)phenyl)methanamine (325 mg, 1.58 mmol, 1 equiv)was added. Upon stirring for 15 h the reaction was diluted with 25 mLEtOAc, and washed with 25 mL brine (2×), followed by 25 mL Sat. NaHCO₃(2×). The organic layer was concentrated down to yield 650 mg (98%yield) of the product as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ 8.67(s, 1H), 7.43-7.29 (m, 4H), 4.49 (d, J=16.7 Hz, 4H), 3.51 (dd, J=11.0,4.7 Hz, 2H), 2.61-2.45 (m, 4H), 2.03 (d, J=7.4 Hz, 2H), 1.42 (s, 9H).TLC: (9:1 DCM:MeOH (0.5 N NH₃)) R_(f)=0.20; MS (ESI) 417.5 (M+H)+.

(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

To (2S,4R)-tert-butyl4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate (650 mg, 1.40 mmol, 1 equiv) in a round bottomflask was charged 9 mL 4M HCL in dioxanes (36 mmol, 26 equiv). Thereaction was left to stir for 1 h upon which time N₂ was bubbled throughfor 1 h and the volatiles were removed by vacuum. The resulting viscousoil was washed dissolved in water and washed with 50 mL EtOAC. Theaqueous layer was then basified to pH 12 with 1 M NaOH, and thenextracted with 50 mL EtOAC (3×). The organic layer was dried andconcentrated down to yield 250 mg (79% yield) of product as a brownviscous oil. 1H NMR (501 MHz, CDCl₃) δ 8.66 (s, 1H), 8.18 (t, J=6.0,1H), 7.38 (d, J=8.1, 2H), 7.30 (d, J=8.1, 2H), 4.48-4.37 (m, 3H), 4.08(t, J=8.4, 1H), 3.02 (d, J=13.3, 1H), 2.79 (dd, J=3.2, 12.3, 1H), 2.51(s, 3H), 2.33 (dd, J=8.6, 13.9, 1H), 2.03-1.87 (m, 1H); ¹³C NMR (126MHz, CDCl₃) δ 174.89, 150.35, 148.46, 138.30, 131.54, 130.95, 129.51,127.92, 72.90, 59.72, 55.35, 42.53, 39.98, 16.06; TLC: (9:1 DCM:MeOH)R_(f)=0.1; LRMS (ESI) 317.4 (M+H)⁺.

General Method E: Representative Procedure: VL219

3-ethoxybenzoic acid (17 mg, 0.1 mmol, 1 equiv.) was dissolved in 1 mL10:1 DCM:DMF and charged with EDC (25 mg, 0.13 mmol 1.3 equiv.), andHOBt (21 mg, 0.13 mmol, 1.3 equiv). After 5 minutes of stirring(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(31 mg, 0.095 mmol, 1 equiv) was added. Upon stirring for 18 h thereaction was diluted with 15 mL EtOAc and washed with 25 mL 10% aqueouscitric acid and 25 mL saturated NaHCO₃. The organic layer was dried withNa₂SO₄ and concentrated by vacuum. The resultant oil was purified bysilica gel chromatography (DCM to 9% MeOH (0.5 N NH₃) in DCM) to yield25 mg (56% yield) of the product as a white solid. ¹H NMR (501 MHz,CD₃OD) δ 8.87 (s, 1H), 7.51-7.42 (m, 4H), 7.37 (t, J=8.1, 1H), 7.23-7.14(m, 2H), 7.05 (dd, J=2.2, 8.4, 1H), 4.79 (dd, J=7.7, 9.5, 1H), 4.63-4.40(m, 3H), 4.08 (q, J=7.0, 2H), 3.86 (dt, J=3.8, 7.6, 1H), 3.47 (d,J=11.5, 1H), 2.47 (s, 3H) 2.36 (dd, J=7.6, 13.2, 1H), 2.14 (ddd, J=5.3,10.2, 16.4, 1H), 1.41 (t, J=7.0, 3H); ¹³C NMR (126 MHz, CD₃OD) δ 174.74,172.64, 160.34, 152.78, 149.05, 140.21, 138.40, 133.39, 131.55, 130.65,130.44, 128.83, 120.49, 118.07, 114.32, 71.02, 64.71, 60.83, 59.81,43.67, 39.30, 15.79, 15.06; TLC: (9:1 DCM:MeOH (0.5 N NH₃)) R_(f)=0.25;LRMS (ESI) 466.1 (M+H)⁺.

VL210 was synthesized according to General Method D. 1H NMR (500 MHz,CD₃OD) δ 8.23 (s, 1H), 7.82 (t, J=1.7, 1H), 7.73-7.58 (m, 4H), 7.49-7.33(m, 4H), 4.76 (dd, J=7.6, 9.6, 1H), 4.59-4.32 (m, 3H), 3.84 (dd, J=3.5,11.4, 1H), 3.41 (d, J=11.3, 1H), 2.43-2.30 (m, 1H), 2.18-2.07 (m, 1H);¹³C NMR (126 MHz, CD₃OD) δ 173.14, 169.54, 151.35, 139.41, 137.87,133.28, 130.18, 130.01, 127.92, 127.61, 126.31, 125.93, 124.23, 121.93,120.36, 69.62, 59.53, 58.30, 42.31, 37.95; LRMS (ESI) 471.5 (M+H)⁺.

VL224 was synthesized according to General Method E. ¹H NMR (501 MHz,CD₃OD) δ 8.87 (s, 1H), 7.83 (d, J=1.5, 1H), 7.67 (d, J=7.1, 1H), 7.61(d, J=6.7, 1H), 7.48-7.36 (m, 5H), 4.77 (t, J=8.5, 1H), 4.60-4.39(obscured m, 3H), 3.90-3.78 (m, 1H), 3.42 (d, J=11.4, 1H), 2.47 (s, 3H),2.41-2.30 (m, 1H), 2.19-2.06 (m, 1H); ¹³C NMR (126 MHz, CD₃OD) δ 173.15,169.55, 151.41, 147.64, 138.77, 137.85, 133.26, 132.78, 131.98, 130.16,130.02, 129.05, 127.43, 125.91, 121.93, 69.64, 59.53, 58.32, 42.27,37.94, 14.41; TLC: (9:1 DCM:MeOH (0.5 N NH₃)) R_(f)=0.7; LRMS (ESI)499.8 (M+H)⁺.

VL215 was synthesized according to General Method D. ¹H NMR (501 MHz,CD₃OD) δ 8.24 (dd, J=13.4, 7.1 Hz, 1H), 7.87-7.58 (m, 3H), 7.58-7.31 (m,4H), 7.16 (s, 1H), 4.73 (d, J=7.8 Hz, 1H), 4.63-4.50 (m, 1H), 4.49-4.29(m, 2H), 3.80 (d, J=10.5 Hz, 1H), 3.60 (s, 1H), 2.31 (s, 1H), 2.17 (s,1H). ¹³C NMR (126 MHz, CD₃OD) δ 179.37, 169.61, 153.12, 152.75, 140.72,138.21, 137.16, 133.72, 130.38, 129.70, 129.40, 129.08, 127.76, 125.63,121.80, 70.64, 60.46, 58.42, 43.80, 39.27. MS (ESI) 504.2 (M+H).

VL228 was synthesized according to General Method E. NMR (400 MHz,CD₃OD) δ 8.88 (d, J=9.3 Hz, 1H), 7.75 (d, J=1.9 Hz, 1H), 7.48 (ddd,J=13.9, 8.4, 4.1 Hz, 5H), 7.17 (d, J=8.3 Hz, 1H), 4.74 (t, J=8.2 Hz,1H), 4.63-4.33 (m, 3H), 3.61 (dd, J=11.3, 3.8 Hz, 1H), 3.18 (d, J=11.3Hz, 1H), 2.49 (d, J=9.6 Hz, 3H), 2.33 (ddd, J=7.8, 4.8, 2.1 Hz, 1H),2.19 (ddd, J=11.9, 7.8, 4.6 Hz, 1H). ¹³C NMR (126 MHz, CD₃OD) δ 173.93,169.45, 152.84, 149.07, 140.11, 138.21, 137.17, 133.73, 131.62, 130.53,130.48, 130.39, 129.52, 129.41, 128.93, 70.65, 60.48, 58.39, 43.75,39.26, 15.81. MS (ESI) 534.4 (M+H).

VL177 was synthesized according to General Method D. ¹H NMR (500 MHz,CDCl₃/CD₃OD) δ 7.89 (s, 1H); 7.84 (s, 1H); 7.80-7.75 (m, 2H); 7.69 (d,J=7.8 Hz, 1H); 7.57-7.48 (m, 3H); 7.31 (d, J=8.1 Hz, 2H); 4.76 (t, J=8.3Hz, 1H); 4.49-4.39 (m, 3H); 3.72 (dd, J=11.2, 3.5 Hz, 1H); 3.36 (d,J=11.2 Hz, 1H); 2.93 (s, 1H); 2.31 (ddd, J=13.2, 8.8, 4.4 Hz, 1H); 2.21(dd, J=13.5, 7.8 Hz, 1H). ¹³C NMR (125 MHz, CDCl₃/CD₃OD) δ 171.8, 169.0,151.5, 150.6, 138.9, 137.0, 133.9, 131.9, 131.3, 129.5, 128.1, 126.7,124.7, 121.1, 117.9, 112.7, 69.9, 59.3, 58.5, 43.3, 37.4. TLC (10% MeOHin CH₂Cl₂), R_(f) 0.17 (UV, CAM), MS (ESI+): calculated for C₂₃H₂₁N₄O₄[M+H]⁺ 417.2, found 417.1.

VL226 was synthesized according to General Method E. ¹H NMR (500 MHz,CDCl₃/CD₃OD) δ 8.65 (s, 1H); 7.84 (s, 1H); 7.74 (dd, J=13.3, 7.8 Hz,2H); 7.53 (t, J=7.8 Hz, 1H); 7.40-7.28 (m, 5H); 4.92 (t, J=8.1 Hz, 1H);4.53 (s, 1H); 4.48 (d, J=5.9 Hz, 2H); 3.72 (dd, J=11.3, 3.5 Hz, 1H);3.52-3.44 (m, 1H); 2.85 (br s, 1H); 2.67-2.56 (m, 1H); 2.48 (s, 3H);2.21 (dd, J=13.5, 7.8 Hz, 1H). ¹³C NMR (125 MHz, CDCl₃/CD₃OD) δ 170.8,169.4, 150.5, 148.6, 138.0, 137.0, 134.1, 131.9, 131.4, 129.7, 129.6,127.0, 118.0, 113.0, 70.4, 59.1, 58.6, 43.5, 36.8, 16.2. TLC (10% MeOHin CH₂Cl₂), R_(f) 0.32 (UV, CAM), MS (ESI+): calculated for C₂₄H₂₃N₄O₃S[M+H]⁺ 447.2, found 447.0.

VL211 was synthesized according to General Method D. 1H NMR (500 MHz,CD₃OD) δ 8.11 (s, 1H), 7.58 (dd, J=2.3, 8.2, 2H), 7.42-7.30 (m, 3H),7.16 (t, J=7.9, 1H), 6.96 (d, J=7.7, 1H), 6.92 (s, 1H), 6.80 (dd, J=2.3,8.1, 1H), 4.65 (t, J=8.6, 1H), 4.47-4.26 (m, 3H), 3.71 (dt, J=4.0, 8.0,1H), 3.36 (d, J=11.6, 1H), 2.32-2.16 (m, 1H), 2.08-1.94 (m, 1H); ¹³C NMR(126 MHz, CD₃OD) δ 174.74, 172.77, 158.71, 153.14, 152.70, 140.83,138.38, 130.62, 128.98, 127.69, 125.62, 121.75, 119.39, 118.61, 115.27,71.01, 60.77, 59.79, 43.73, 39.25; TLC: (9:1 DCM:MeOH R=0.15; LRMS (ESI)408.3 (M+H)+.

Vl225 was synthesized according to General Method E. ¹H NMR (501 MHz,CD₃OD) δ 8.86 (s, 1H), 7.49-7.34 (m, 4H), 7.27 (t, J=7.8, 1H), 7.11-7.00(m, 2H), 6.93-6.84 (m, 1H), 4.77 (t, J=8.5, 1H), 4.57-4.38 (m, 3H), 3.84(dd, J=3.3, 11.5, 1H), 3.47 (d, J=11.5, 1H), 2.46 (S, 3H) 2.34 (dd,J=7.1, 12.5, 1H), 2.18-2.06 (m, 1H); ¹³C NMR (126 MHz, CD₃OD) δ 173.35,171.36, 157.31, 151.39, 147.62, 138.81, 136.95, 130.12, 129.21, 129.04,127.62, 127.41, 117.97, 117.19, 113.84, 69.61, 59.37, 58.41, 42.24,37.85, 14.37; TLC: (9:1 DCM:MeOH) R=0.3; LRMS (ESI) 437.0 (M+H)⁺.

VL178 was synthesized according to General Method D. ¹H NMR (400 MHz,CD₃OD) δ 8.84 (t, J=5.9, 1H), 8.26 (d, J=5.9, 1H), 7.73 (d, J=8.3, 2H),7.55-7.45 (m, 3H), 7.06 (t, J=7.8, 1H), 6.86-6.76 (m, 1H), 6.68 (d,J=7.3, 1H), 4.75 (t, J=8.4, 1H), 4.66-4.35 (m, 3H), 3.55 (dd, J=3.5,11.6, 1H), 3.24 (d, J=11.4, 1H), 2.41-2.26 (m, 1H), 2.21-2.10 (m, 4H);¹³C NMR (126 MHz, CD₃OD) δ 173.21, 172.03, 151.32, 145.03, 139.46,137.30, 130.95, 127.61, 126.53, 126.29, 124.22, 123.96, 120.35, 116.31,115.97, 74.46, 69.35, 58.72, 42.46, 38.02, 23.61; LRMS (ESI) 420.4(M+H)⁺.

(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(24 mg, 0.0756 mmol, 1 eq), 3-amino-2-methylbenzoic acid (13 mg, 0.083mmol, 1.1 eq), EDC (16 mg, 0.083 mmol, 1.1 eq) and HOBt (11 mg, 0.083mmol, 1.1 eq) were dissolved in DMF (0.76 mL) at room temperature. DIPEA(0.02 mL, 0.113 mmol, 1.5 eq) was added, and the solution was stirredfor 17 hours. The solution was then partitioned between 1M NaOH andEtOAc, separated, and extracted twice more with EtOAc. The combinedorganic layer was dried over sodium sulfate, filtered and condensed.Purification by column chromatography (1 to 10% 0.5N methanolicammonia/DCM) gave a white solid (20.5 mg, 0.045 mmol, 60%). ¹H NMR (501MHz, CD₃OD) δ 8.87 (t, J=6.6 Hz, 1H), 7.45 (dt, J=20.5, 7.7 Hz, 4H),7.03 (t, J=7.6 Hz, 1H), 6.78 (d, J=7.8 Hz, 1H), 6.65 (s, 1H), 4.74 (t,J=8.1 Hz, 1H), 4.67-4.34 (m, 3H), 3.53 (d, J=11.3 Hz, 1H), 3.21 (d,J=9.3 Hz, 1H), 2.48 (d, J=3.1 Hz, 3H), 2.32 (d, J=7.6 Hz, 1H), 2.14 (dd,J=31.4, 20.5 Hz, 4H). ¹³C NMR (126 MHz, CD₃OD) δ 174.56, 173.67, 152.89,152.81, 149.04, 147.80, 140.23, 138.57, 133.44, 131.54, 130.46, 128.86,127.83, 117.01, 116.36, 70.77, 69.66, 60.09, 43.70, 39.41, 15.81, 13.92.MS (ESI) 450.6 (M+H), 473.4 (M+Na).

For further reference see the following articles and the referencescited therein:

-   (1) Buckley D L et al. J. Am. Chem. Soc 2012, 134, 4465-4468.-   (2) Van Molle I et al. A Chemistry & Biology 2012, 19, 1300-1312-   (3) Buckley, D Angew. Chem. Int. Ed., 2012, 51, 11463-11467-   (4) Buckley, D. L et al. Angew. Chem. 2012, 124, 11630-11634.

Examples—Compounds 165-266 of Affinity Table II

The following compounds were synthesized according to the stated GeneralMethod, were purified by standard chromatographic methods and had ¹H and¹³C NMR and MS data consistent with the desired structure.

VL165 was synthesized according to General Method B.

VL168 & 169

The chiral RHS amine fragment was synthesized using the procedure fromSurya Prakash, G. K.; Mandal, M.; Olah, G. A. Angew. Chem. Int. Ed.2001, 40, 589-690.

VL168 was Synthesized According to General Method F

VL169 was Synthesized According to General Method F

VL175: General Method C

VL192-VL205: Solid Phase Synthesis General Method B

3-hydroxy-2-methylbenzoic acid (26.3 mg, 0.173 mmol, 1.1 eq), EDC (33.2mg) and HOBt (23.4 mg) were dissolved in DCM (0.8 mL) and DMF (0.1 mL)at 4° C. After 10 minutes,(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(0.5 mL of a 100 mg/mL solution in DCM) was added and the mixture waswarmed to room temperature. After 21 hours, the mixture was diluted with10 mL of half saturated sodium chloride and extracted thrice with 10 mLof EtOAc. The combined organic layer was dried over sodium sulfate,filtered and condensed. Purification by column chromatography (1 to 10%MeOH/DCM) gave a white solid (29.2 mg, 0.0647, 41%). ¹H NMR (400 MHz,MeOD) δ 8.93-8.82 (m, 1H), 7.55-7.36 (m, 4H), 7.10 (t, J=7.8 Hz, 1H),6.83 (dd, J=10.3, 7.9 Hz, 2H), 4.76 (t, J=8.4 Hz, 1H), 4.66-4.38 (m,3H), 3.56 (dd, J=11.6, 3.6 Hz, 1H), 3.22 (d, J=11.6 Hz, 1H), 2.54-2.47(m, 3H), 2.41-2.31 (m, 1H), 2.19 (dt, J=11.3, 10.9 Hz, 4H). ¹³C NMR (126MHz, MeOD) δ 174.50, 173.09, 157.22, 152.89, 152.80, 149.01, 140.21,139.18, 133.42, 131.52, 130.44, 128.85, 127.94, 117.84, 116.42, 70.73,69.62, 60.12, 43.69, 39.38, 15.79, 12.70. MS (ESI) 452.5 (M+H).

VL251(2S,4R)-1-((S)-2-((S)-2-acetamido-4-methylpentanamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

tert-butyl((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-yl)carbamate

Boc-Ala-OH (189 mg, 1.0-mmol) was dissolved in 10 mL DCM and chargedwith EDC (248 mg, 1.2 mmol), and HOBt (202 mg, 1.3 mmol) after 5 minutesof stirring(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carb-oxamide(317 mg, 1.0 mmol) was added. Upon stirring for 18 h the reaction wasdiluted with 10 mL DCM and washed with 10 mL 10% aqueous citric acidfollowed by 5 mL saturated NaHCO₃. The mixture was concentrated down andpurified by silica gel chromatography (DCM/MeOH gradient) to yield 210mg (43% yield) of the product as a white solid. 1H NMR (501 MHz, CDCl₃)δ 8.65 (s, 1H), 7.58 (s, 1H), 7.31 (d, J=8.1, 2H), 7.26 (d, J=8.0, 2H),5.44 (d, J=7.4, 1H), 4.66 (t, J=7.6, 1H), 4.52 (s, 1H), 4.39 (m, 3H),3.78 (d, J=10.9, 1H), 3.59 (d, J=7.0, 1H), 2.47 (s, 3H), 2.30 (s, 1H),2.10 (s, 1H), 1.54-1.31 (m, 9H), 1.26 (d, J=6.9, 3H); ¹³C NMR (126 MHz,CDCl₃) δ 173.1, 171.2, 155.5, 150.5, 148.2, 138.2, 130.7, 129.4, 127.6,80.1, 70.1, 58.8, 55.2, 48.0, 42.3, 36.5, 28.3, 18.0, 16.0; LRMS (ESI)489.4 (M+H)⁺.

VL251(2S,4R)-1-((S)-2-((S)-2-acetamido-4-methylpentanamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

Boc-Ala-Hyp-benzyl thiazole (116 mg, 0.225 mmol) was dissolved in 1 mLDCM and charged with 2.3 mL 4M HCL in dioxanes. Upon stirring for onehour Nitrogen gas was sparged through the mixture for 15 minutes and theremaining volatiles removed by roto vap.

The resultant foam was then dissolved in 5 mL 1:1 DCM: DMF and chargedwith EDC (56 mg, 0.29 mmol), HOBt (45 mg, 0.29 mmol), and Ac-Leu-OH (43mg, 0.25 mmol) were added. After stirring for 5 minutes triethyl aminewas added. Upon stirring for 18 h the reaction was diluted with 10 mLEtOAc and washed with 10 mL 10% aqueous citric acid followed by 5 mLsaturated NaHCO₃ The aqueous layer was then back extracted 2×10 mL DCM.The organic layers were combined and the mixture was concentrated downand purified by silica gel chromatography (DCM/MeOH gradient) to yield35 mg (29% yield) of the product as a white solid. 1H NMR (501 MHz,CDCl₃) δ 8.68 (s, 1H), 8.05 (s, 1H), 7.70 (s, 1H), 7.36 (d, J=7.3, 2H),7.28 (d, J=8.6, 2H), 6.50 (s, 1H), 4.85-4.73 (m, 2H), 4.68 (s, 1H), 4.59(s, 1H), 4.40 (d, J=32.4, 2H), 3.84 (d, J=10.7, 1H), 3.70 (d, J=10.5,1H), 2.51 (s, 3H), 2.30 (s, 1H), 2.21 (s, 1H), 1.85 (s, 3H), 1.59 (s,1H), 1.50 (s, 2H), 1.34 (d, J=6.7, 3H), 0.86 (t, J=6.7, 6H); ¹³C NMR(126 MHz, CDCl₃) δ 172.1, 172.0, 171.2, 170.8, 150.4, 148.4, 138.1,129.5, 129.4, 127.8, 110.0, 70.36, 58.9, 55.5, 51.8, 46.9, 43.1, 41.8,36.9, 24.7, 23.2, 23.1, 21.8, 17.9, 16.0; LRMS (ESI) 545.1 (M+H)⁺.

VL252(2S,4R)-1-((S)-2-((S)-2-amino-4-methylpentanamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

Boc-Ala-Hyp-benzyl thiazole (116 mg, 0.225 mmol) was dissolved in 1 mLDCM and charged with 2.3 mL 4M HCL in dioxanes. Upon stirring for onehour Nitrogen gas was sparged through the mixture for 15 minutes and theremaining volatiles removed by roto vap. The resultant foam was thendissolved in 5 mL 1:1 DCM: DMF and charged with EDC (56 mg, 0.29 mmol),HOBt (45 mg, 0.29 mmol), and Boc-Leu-OH (62 mg, 0.25 mmol) were added.After stirring for 5 minutes triethyl amine was added. Upon stirring for18 h the reaction was diluted with 10 mL EtOAc and washed with 10 mL 10%aqueous citric acid followed by 5 mL saturated NaHCO₃. The organiclayers were combined and the mixture was concentrated to yield 55 mg(40% yield) of the product as a white solid. LRMS (ESI) 602.0 (M+H)+.Upon confirmation by mass spec the product was dissolved in 2 mL 1:1DCM:MeOH and charged with 3 mL 4M HCl in dioxanes. Upon stirring for 45minutes the reaction was quenched with 5 ml 0.5 N ammonia in methanol.The solvents were evaporated down and purified by silica gelchromatography (gradient of DCM/MeOH (0.5 N NH₃) to yield 50 mg of pureproduct as a white solid. 1H NMR (501 MHz, CDCl₃) δ 8.25 (s, 1H), 6.91(dd, J=7.3, 18.0, 4H), 4.17 (d, J=7.5, 2H), 4.06 (d, J=22.4, 2H), 3.95(d, J=15.3, 1H), 3.56-3.42 (m, 2H), 3.24 (d, J=7.8, 1H), 2.05 (s, 3H),1.86-1.77 (m, 1H), 1.77-1.65 (m, 1H), 1.35-1.19 (m, 2H), 1.13 (s, 1H),0.93 (d, J=6.8, 3H), 0.50 (dd, J=6.3, 9.8, 6H); ¹³C NMR (126 MHz, CDCl₃)δ 171.6, 171.5, 150.3, 147.7, 137.9, 131.4, 130.1, 129.0, 127.3, 109.9,69.7, 58.6, 55.11, 46.9, 42.5, 36.7, 24.1, 22.4, 22.2, 16.2, 15.3; LRMS(ESI) 502.0 (M+H)⁺.

Examples for Compounds of FIG. 15

The following procedures were used to synthesize and/or characterizecompounds according to the present invention as indicated

LCMS Method:

The analysis was conducted on an Acquity UPLC BEH C18 column (50 mm×2.1mm internal diameter 1.7 μm packing diameter) at 40° C.

The solvents employed were:

A=0.1% v/v solution of formic acid in water.B=0.1% v/v solution of formic acid in acetonitrile.

The gradient employed was as follows:

Time Flow Rate (minutes) (mL/min) % A % B 0 1 97 3 1.5 1 0 100 1.9 1 0100 2.0 1 97 3

The UV detection was an averaged signal from wavelength of 210 nm to 350nm and mass spectra were recorded on a mass spectrometer usingalternate-scan positive and negative mode electrospray ionization.

The following illustrates the mobile phases and gradients used whencompounds underwent purification by mass-directed autopreparative HPLC.

Mass-Directed Autopreparative HPLC (Formic Acid Modifier)

The HPLC analysis was conducted on a Sunfire C18 column (150 mm×30 mminternal diameter, 5 m packing diameter) at ambient temperature.

The solvents employed were:

A=0.1% v/v solution of formic acid in water.B=0.1% v/v solution of formic acid in acetonitrile.

Mass-Directed Autopreparative HPLC (Trifluoroacetic Acid Modifier)

The HPLC analysis was conducted on a Sunfire C18 column (150 mm×30 mminternal diameter, 5 m packing diameter) at ambient temperature.

The solvents employed were:

A=0.1% v/v solution of trifluoroacetic acid in water.B=0.1% v/v solution of trifluoroacetic acid in acetonitrile.

Mass-Directed Autopreparative HPLC (Ammonium Bicarbonate Modifier) TheHPLC analysis was conducted on an XBridge C18 column (150 mm×30 mminternal diameter, 5 m packing diameter) at ambient temperature.

The solvents employed were:

A=10 mM ammonium bicarbonate in water adjusted to pH 10 with ammoniasolution.B=acetonitrile.

For each of the mass-directed autopreparative purifications,irrespective of the modifier used, the gradient employed was dependentupon the retention time of the particular compound undergoingpurification as recorded in the analytical LCMS, and was as follows:

For compounds with an analytical LCMS retention time below 0.6 minutesthe following gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B 0 40 99 1 1 40 99 1 10 40 7030 11 40 1 99 15 40 1 99

For compounds with an analytical LCMS retention time between 0.6 and 0.9minutes the following gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B 0 40 85 15 1 40 85 15 10 40 4555 11 40 1 99 15 40 1 99

For compounds with an analytical LCMS retention time between 0.9 and 1.2minutes the following gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B 0 40 70 30 1 40 70 30 10 40 1585 11 40 1 99 15 40 1 99

For compounds with an analytical LCMS retention time between 1.2 and 1.4minutes the following gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B 0 40 50 50 1 40 50 50 10 40 199 11 40 1 99 15 40 1 99

The UV detection was an averaged signal from wavelength of 210 nm to 350nm and mass spectra were recorded on a mass spectrometer usingalternate-scan positive and negative mode electrospray ionization.

The chemical names were generated using ACD Name Pro version 6.02 fromAdvanced Chemistry Development, Inc.

EXAMPLES(2S,4R)-1-(2-ethoxybenzoyl)-4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A solution of 2-ethoxybenzoic acid (commercially available from forexample Aldrich) (29 mg, 0.17 mmol),(2S,4R)-4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide (50mg, 0.17 mmol) and DIPEA (0.061 mL, 0.35 mmol) in DMF (1 mL) was treatedwith HATU (80 mg, 0.21 mmol) and the mixture was stirred at ambienttemperature for 2 hours. The product was then subjected to purificationby mass-directed automated preparative HPLC (formic acid modifier) toafford the title compound (38 mg, 0.087 mmol, 50% yield). LCMS RT=0.72min, ES+ve m/z 436 [M+H]⁺.

Using a method analogous to that for(2S,4R)-1-(2-ethoxybenzoyl)-4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide,the following compounds were prepared:

Name Structure Yield RT [M + H]+ (2S,4R)-1-benzoyl-4-hydroxy-N-(4-(oxazol-5- yl)benzyl)pyrrolidine-2- carboxamide

82% 0.65 min 392 (2S,4R)-4-hydroxy-1-(3- methoxybenzoyl)-N-(4-(oxazol-5- yl)benzyl)pyrrolidine-2- carboxamide

66% 0.67 min 422 (2S,4R)-1-(3-ethoxybenzoyl)- 4-hydroxy-N-(4-(-oxazol-5-yl)benzyl)pyrrolidine-2- carboxamide

54% 0.72 min 436 (2S,4R)-1-(4-ethoxybenzoyl)- 4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2- carboxamide

46% 0.71 min 436 (2S,4R)-1-(3-cyanobenzoyl)- 4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2- carboxamide

57% 0.64 min 417 (2S,4R)-4-hydroxy-1-(3- isopropoxybenzoyl)-N-(4-(oxazol-5- yl)benzyl)pyrrolidine-2- carboxamide

53% 0.78 min 450 (2S,4R)-1-(3-acetylbenzoyl)- 4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2- carboxamide

47% 0.63 min 434 (2S,4R)-4-hydroxy-1-(3- morpholinobenzoyl)-N-(4-(oxazol-5- yl)benzyl)pyrrolidine-2- carboxamide

63% 0.66 min 477 (2S,4R)-4-hydroxy-1-(3- isopropylbenzoyl)-N-(4-(oxazol-5- yl)benzyl)pyrrolidine-2- carboxamide

69% 0.82 min 434 (2S,4R)-1-(3-chlorobenzoyl)- 4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2- carboxamide

65% 0.73 min 426 (2S,4R)-1-(3-bromobenzoyl)- 4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2- carboxamide

66% 0.75 min 470, 472 (2S,4R)-1-(3-ethylbenzoyl)-4-hydroxy-N-(4-(oxazol-5- yl)benzyl)pyrrolidine-2- carboxamide

55% 0.77 min 420 (2S,4R)-1-(3,5- diethoxybenzoyl)-4-hydroxy-N-(4-(oxazol-5- yl)benzyl)pyrrolidine-2- carboxamide

65% 0.81 min 480 (2S,4R)-4-hydroxy-N-(4- (oxazol-5-yl)benzyl)-1-(3-propoxybenzoyl)pyrrolidine- 2-carboxamide

52% 0.80 min 450

(S)-1-((2S,4R)-4-hydroxy-2-((4-(oxazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-ylacetate and(2S,4R)-4-hydroxy-1-((S)-2-hydroxypropanoyl)-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamidehydrochloride (60 mg, 0.19 mmol) and (S)-2-acetoxypropanoic acid(commercially available from for example Aldrich) (25 mg, 0.19 mmol) inDMF (1.2 mL) was treated with DIPEA (0.13 mL, 0.74 mmol) and then withHATU (85 mg, 0.22 mmol) and the mixture was stirred at ambienttemperature for 30 minutes. Half of the reaction mixture was thensubjected to purification by mass-directed automated preparative HPLC(formic acid modifier) to afford (S)-1-((2S,4R)-4-hydroxy-2-((4-(oxazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-ylacetate (21 mg, 0.052 mmol, 28% yield). LCMS RT=0.58 min, ES+ve m/z 402[M+H]⁺.

The remaining half of the reaction mixture was treated with ammonia (2Msolution in methanol) (2 mL), sealed and allowed to stand for 1 day. Thesolution was then evaporated to dryness and the product was subjected topurification by mass-directed automated preparative HPLC (formic acidmodifier) to afford(2S,4R)-4-hydroxy-1-((S)-2-hydroxypropanoyl)-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide(18 mg, 0.050 mmol, 27% yield). LCMS RT=0.53 min, ES+ve m/z 360 [M+H]⁺.

(2S,4R)-benzyl4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylate

An ice-cooled mixture of 2-(3-methylisoxazol-5-yl)acetic acid(commercially available from for example Aldrich) (0.91 g, 6.4 mmol) and(2S,4R)-benzyl 4-hydroxypyrrolidine-2-carboxylate, hydrochloride (1.67g, 6.5 mmol) in DMF (9 mL) was treated with DIPEA (3.4 mL, 19 mmol) andthen with HATU (2.56 g, 6.7 mmol) over 20 minutes. The mixture wasstirred with cooling for 30 minutes and then overnight at ambienttemperature. The mixture was then treated with saturated aqueous sodiumbicarbonate (50 mL), extracted with dichloromethane (4×60 mL) and thecombined organic phase was filtered through a hydrophobic frit andevaporated to dryness. The product was purified by flash chromatography(100 g cartridge) using a gradient elution from 0% to 15% methanol indichloromethane to afford the title compound (2.3 g, 6.7 mmol,quantitative). LCMS RT=0.75 min, ES+ve m/z 345 [M+H]⁺.

(2S,4R)—N-((3,4-dihydro-2H-benzo[b][1,4]oxazin-2-yl)methyl)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide

A solution of(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid (90 mg, 0.35 mmol),(3,4-dihydro-2H-benzo[b][1,4]oxazin-2-yl)methanamine (commerciallyavailable from for example Fluorochem) (58 mg, 0.35 mmol) and DIPEA(0.155 mL, 0.89 mmol) in DMF (2 mL) was treated with HATU (139 mg, 0.37mmol) and stirred for 1 hour. The product was subjected to purificationby mass-directed automated preparative HPLC (formic acid modifier) toafford the title compound (84 mg, 0.21 mmol, 60% yield) LCMS RT=0.61min, ES+ve m/z 401 [M+H]⁺.

(2S,4R)—N-(4-chlorobenzyl)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide

A solution of (4-chlorophenyl)methanamine (commercially available fromfor example Aldrich) (0.021 mL, 0.17 mmol) and(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid (40 mg, 0.16 mmol) in DMF (1 mL) was treated with DIPEA (0.082 mL,0.47 mmol) then with HATU (66 mg, 0.17 mmol) and the mixture was stirredat ambient temperature for 2 hours. The product was subjected topurification by mass-directed automated preparative HPLC (formic acidmodifier) to afford the title compound (24 mg, 0.064 mmol, 40% yield).LCMS RT=0.73 min, ES+ve m/z 378 [M+H]⁺.

(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)-N-(4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid (60 mg, 0.24 mmol) and5-(4-(aminomethyl)phenyl)benzo[d]oxazol-2(3H)-one, hydrochloride (65 mg,0.24 mmol) in DMF (1.6 mL) was treated with DIPEA (0.124 mL, 0.71 mmol)and HATU (99 mg, 0.26 mmol) and the mixture was stirred for 30 minutes.The product was then subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (64 mg, 0.13 mmol, 57% yield). LCMS RT=0.70 min, ES+ve m/z 477[M+H]⁺.

(2S,4R)—N-(1-(benzofuran-2-yl)ethyl)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide

A stirred solution of(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid (90 mg, 0.35 mmol), 1-(benzofuran-2-yl)ethanamine (commerciallyavailable from for example Enamine) (57 mg, 0.35 mmol) and DIPEA (0.155mL, 0.89 mmol) in DMF (2 mL) was treated with HATU (139 mg, 0.37 mmol).After 1 hour the product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (91 mg, 0.21 mmol, 65% yield) LCMS RT=0.79 min, ES+ve m/z 398[M+H]⁺.

(2S,4R)—N-([1,1′-biphenyl]-4-ylmethyl)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid (30 mg, 0.12 mmol) and [1,1′-biphenyl]-4-ylmethanamine(commercially available from for example Aldrich) (22 mg, 0.12 mmol) inDMF (0.8 mL) was treated with DIPEA (0.08 mL, 0.47 mmol) and then withHATU (49 mg, 0.13 mmol) and the mixture was stirred at ambienttemperature for 30 minutes. The product was subjected to purification bymass-directed automated preparative HPLC (formic acid modifier) toafford the title compound (40 mg, 81% yield). LCMS RT=0.86 min, ES+vem/z 420 [M+H]⁺.

Using a method analogous to that for(2S,4R)—N-([1,1′-biphenyl]-4-ylmethyl)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamidethe following compounds were prepared:

Name Structure Yield RT [M + H]+ (2S,4R)-N- benzyl-4- hydroxy-1-(2-(3-methylisoxazol-5- yl)acetyl)pyrrolidine- 2-carboxamide

61% 0.62 min 344 (2S,4R)-4- hydroxy-N-((1- methyl-1H- pyrazol-3-yl)methyl)-1-(2- (3- methylisoxazol-5- yl)acetyl)pyrrolidine-2-carboxamide

75% 0.40 min 348 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N-((6- phenylpyridin-3- yl)methyl)pyrro- lidine-2-carboxamide

41% 0.57 min 421 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N-((2- oxo-2,3-dihydro- 1H- benzo[d]imidazol- 5-yl)methyl)pyrro- lidine-2- carboxamide

24% 0.42 min 400 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N- (quinolin-7- ylmethyl)pyrrolidine- 2-carboxamide

19% 0.39 min 395 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N-((4- phenylthiazol-2- yl)methyl)pyrro- lidine-2-carboxamide

34% 0.72 min 427 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N-((1- phenylpyrrolidin- 3- yl)methyl)pyrro- lidine-2-carboxamide

35 0.72 min 413 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N-((5- phenyl-1,2,4- oxadiazol-3- yl)methyl)pyrro- lidine-2-carboxamide

31% 0.67 min 412 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N-((3- (p-tolyl)isoxazol- 5- yl)methyl)pyrro- lidine-2-carboxamide

56% 0.78 min 426 (2S,4R)-N-((S)-1- (4- chlorophenyl)eth-yl)-4-hydroxy-1- (2-(3- methylisoxazol-5- yl)acetyl)pyrrolidine-2-carboxamide

58% 0.78 min 392 (2S,4R)-N-((3-(4- chlorophenyl)- 1,2,4-oxadiazol-5-yl)methyl)-4- hydroxy-1-(2-(3- methylisoxazol-5- yl)acetyl)pyrrolidine-2-carboxamide

57% 0.81 min 446 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N-((5- phenylisoxazol-3- yl)methyl)pyrro- lidine-2-carboxamide

66% 0.72 min 411 (2S,4R)-4- hydroxy-1-(2-(3- methylisoxazol-5-yl)acetyl)-N-(4- (pyrrolidin-1- yl)benzyl)pyrro- lidine-2- carboxamide

43% 0.60 min 413

(2S,4R)-1-((S)-2-acetamidopropanoyl)-4-hydroxy-N-phenethylpyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-1-((S)-2-acetamidopropanoyl)-4-hydroxypyrrolidine-2-carboxylicacid (24 mg, 0.10 mmol) and 2-phenylethanamine (commercially availablefrom for example Aldrich) (0.012 mL, 0.10 mmol) in DMF (0.8 mL) wastreated with DIPEA (0.07 mL, 0.39 mmol) and then with HATU (45 mg, 0.12mmol), and the mixture was stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (25 mg, 72% yield). LCMS RT=0.56 min, ES+ve m/z 348 [M+H]⁺.

Using a method analogous to that for(2S,4R)-1-((S)-2-acetamidopropanoyl)-4-hydroxy-N-phenethylpyrrolidine-2-carboxamidethe following compounds were prepared:

[M + Name Structure Yield RT H]+ (2S,4R)-1-((S)-2-acetamidopropanoyl)-4-hydroxy- N-((6-phenylpyridin-3-yl)methyl)pyrrolidine-2- carboxamide

10% 0.49 min 411 (2S,4R)-N-([1,1′-biphenyl]-4- ylmethyl)-1-((S)-2-acetamidopropanoyl)-4- hydroxypyrrolidine-2-carboxamide

40% 0.79 min 411 (2S,4R)-1-((S)-2- acetamidopropanoyl)-N-benzyl-4-hydroxypyrrolidine-2-carboxamide

12% 0.51 min 334 (2S,4R)-1-((S)-2- acetamidopropanoyl)-4-hydroxy-N-((3-(p-tolyl)isoxazol-5- yl)methyl)pyrrolidine-2- carboxamide

70% 0.70 min 415 (2S,4R)-1-((S)-2- acetamidopropanoyl)-N-cinnamyl-4-hydroxypyrrolidine-2- carboxamide

50% 0.64 min 360 (2S,4R)-1-((S)-2- acetamidopropanoyl)-N-(2-(benzyl(methyl)amino)-2- oxoethyl)-4-hydroxypyrrolidine-2- carboxamide

27% 0.56 min 405 (2S,4R)-1-((S)-2- acetamidopropanoyl)-4-hydroxy-N-(2-((3- methoxybenzyl)(methyl)amino)-2- oxoethyl)pyrrolidine-2-carboxamide

28% 0.58 min 435 (2S,4R)-1-((S)-2- acetamidopropanoyl)-4-hydroxy-N-((1S,2R)-2- phenylcyclopropyl)pyrrolidine-2- carboxamide

44% 0.62 min 360 (2S,4R)-1-((S)-2- acetamidopropanoyl)-N-((R)-1-benzyl-2-oxopyrrolidin-3-yl)-4- hydroxypyrrolidine-2-carboxamide

76% 0.58 min 417

(S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-ylacetate &(2S,4R)-1-acetyl-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

An ice-cooled mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (31 mg, 0.088 mmol) and (S)-2-acetoxypropanoic acid(commercially available from for example Aldrich) (8 μL, 0.09 mmol) inDMF (0.8 mL) was treated with DIPEA (0.074 mL, 0.42 mmol). HATU (34 mg,0.088 mmol) was then added portion-wise over 10 minutes and the mixturewas stirred at ambient temperature for 1 hour. The products wereseparated and purified by mass-directed automated preparative HPLC(formic acid modifier) to afford(S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-ylacetate (15 mg, 0.035 mmol, 41% yield) LCMS RT=0.64 min, ES+ve m/z 432[M+H]⁺ and(2S,4R)-1-acetyl-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(9 mg, 0.025 mmol, 30% yield) LCMS RT=0.57 min, ES+ve m/z 360 [M+H]⁺.

(2S,4R)-1-(2-(cyanomethyl)benzoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-4-hydroxy-N-(4-(thiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(50 mg, 0.17 mmol) and 2-(cyanomethyl)benzoic acid (commerciallyavailable from for example Aldrich) (29 mg, 0.18 mmol) in DMF (0.7 mL)was treated with DIPEA (0.086 mL, 0.49 mmol) and then with HATU (69 mg,0.18 mmol) and the mixture was stirred at ambient temperature for 10minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (31 mg, 0.067 mmol, 41% yield). LCMS RT=0.73 min, ES+ve m/z 461[M+H]⁺.

Using a method analogous to that for(2S,4R)-1-(2-(cyanomethyl)benzoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamidethe following compounds were prepared:

Stereo- chemistry [M + Name Structure Comment Yield RT H]+(2S,4R)-4-hydroxy-1-(2-(2- methoxyethoxy)acetyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

59% 0.57 min 434 (2S,4R)-1-((S)-2- acetamidopropanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

71% 0.56 min 431 (2S,4R)-1-((S)-1- acetylpyrrolidine-2-carbonyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

57% 0.54 min 457 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- ((S)-5-oxopyrrolidine-2-carbonyl)pyrrolidine-2- carboxamide

58% 0.53 min 429 (2S,4R)-1- (cyclohexanecarbonyl)-4- hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

61% 0.76 min 428 (2S,4R)-1-(4-ethoxybenzoyl)- 4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

73% 0.78 min 466 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- (5-oxopyrrolidine-3-carbonyl)pyrrolidine-2- carboxamide

85% 0.53 min 429 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- (2- morpholinoacetyl)pyrrolidine-2-carboxamide

40% 0.48 min 445 (2S,4R)-4-hydroxy-1-(2- methoxyacetyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

68% 0.55 min 390 (2S,4R)-4-hydroxy-1-((S)-2- methoxypropanoyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

69% 0.57 min 404 (2S,4R)-4-hydroxy-1-(3- methoxybenzoyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

67% 0.71 min 452 (2S,4R)-4-hydroxy-1-(2- methylbenzoyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

77% 0.74 min 436 (2S,4R)-4-hydroxy-1-(3- methoxy-2-methylbenzoyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

61% 0.76 min 466 (2S,4R)-1-(3-chloro-5- methoxybenzoyl)-4-hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

69% 0.80 min 486 (2S,4R)-4-hydroxy-1-(3- hydroxy-2-methylbenzoyl)-N-(4-(4-methyithiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

84% 0.64 min 452 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- (2-(3- oxomorpholino)propanoyl)pyrrolidine-2-carboxamide

Single enantiomer, stereo- chemistry unknown at the unspecified chiralcentre, eluted first during HPLC purification (formic acid modifier) 32%0.59 min 473 (2S,4R)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1-((S)-2-(1-oxoisoindolin-2- yl)pentanoyl)pyrrolidine-2- carboxaniide

69% 0.84 min 533 (2S,4R)-4-hydroxy-1-(2-(6- methoxy-1-oxoisoindolin-2-yl)-3-methylbutanoyl)-N-(4- (4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

Single enantiomer, stereo- chemistry unknown at the unspecified chiralcentre, eluted first during HPLC purification (formic acid modifier) 35%0.84 min 563 (2S,4R)-4-hydroxy-1-(2-(6- methoxy-1-oxoisoindolin-2-yl)-3-methylbutanoyl)-N-(4- (4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

Single enantiomer, stereo- chemistry unknown at the unspecified chiralcentre, eluted first during HPLC purification (formic acid modifier) 34%0.85 min 563 (2S,4R)-4-hydroxy-1-(2-(5- methoxy-1-oxoisoindolin-2-yl)-3-methylbutanoyl)-N-(4- (4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

Single enantiomer, stereo- chemistry unknown at the unspecified chiralcentre, eluted first during HPLC purification (formic acid modifier) 28%0.84 min 536 (2S,4R)-1-(2-(7-chloro-1- oxoisoindolin-2-yl)-3-methylbutanoyl)-4-hydroxy- N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the unspecified chiralcentre, eluted first during HPLC purification (formic acid modifier) 35%0.93 min 567, 569 (2S,4R)-1-((S)-2-acetamido-4-methylpentanoyl)-4-hydroxy- N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

71% 0.71 min 473 (2S,4R)-1-((S)-2-acetamido-3-phenylpropanoyl)-4-hydroxy- N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

77% 0.75 min 507 (2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(3-methyl-2,5-dioxo- 2,5-dihydro-1H-pyrrol-1-yl)butanoyl)-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

17% 0.80 min 511 (2S,4R)-1-benzoyl-4-hydroxy- N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

57% 0.69 min 422 (2S,4R)-1-(3- (cyanomethyl)benzoyl)-4- hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

74% 0.68 min 461 (2S,4R)-1-(1-(cyanomethyl)- 1H-indole-2-carbonyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

62% 0.82 min 500 (2S,4R)-1-(4- ethoxycyclohexanecarbonyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

74% 0.74 min 472 (2S,4R)-1-(2- cyclopentylpropanoyl)-4- hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

Mixture of diastereo- isomers 67% 0.82 min 442(2S,4R)-4-hvdroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1- (2-morpholinopropanoyl) pyrrolidine-2-carboxamide

Mixture of diastereo- isomers 53% 0.68 min 459 (2S,4R)-4-hydroxy-1-(indoline-2-carbonyl)-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during 64% 0.70 min 463 HPLC purification (formicacid modifier) (2S,4R)-1-(2-(4-chloro-1H- pyrazol-1-yl)propanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC 39% 0.74 min 474 purification (formicacid modifier) (2S,4R)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1-(2-(1-oxoisoindolin-2- yl)propanoyl)pyrrolidine-2- carboxamide

Mixture of diastereo- isomers 51% 0.73 min 505 (2S,4R)-1-(2-(3-(difluoromethyl)-5-methyl- 1H-pyrazol-1-yl)propanoyl)- hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC purification 31% 0.76 min 504 (formicacid modifier) (2S,4R)-4-hydroxy-1-(1H- indole-2-carbonyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

65% 0.78 min 461 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- (2-(pyrazolo[1,5-a]pyridin-3-yl)acetyl)pyrrolidine-2- carboxamide

51% 0.65 min 476 (2S,4R)-1-(2- cyclohexylpropanoyl)-4- hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

Mixture of diastereo- isomers 68% 0.87 min 456 (2S,4R)-1-(2-((4-fluorophenyl)amino)propanoyl)- 4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Mixture of diastereo- isomers 41% 0.77 min 483(2S,4R)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1-(2-(1-oxoisoindolin-2- yl)acetyl)pyrrolidine-2- carboxamide

49% 0.69 min 491 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- (2-(pyrazolo[1,5-a]pyridin-3-yl)propanoyl)pyrrolidine-2- carboxamide

Mixture of diastereo- isomers 61% 0.70 min 490 (2S,4R)-1-(2-(3-cyanophenyl)propanoyl)-4- hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Mixture of diastereo- isomers 54% 0.77 min 475 (2S,4R)-4-hydroxy-1-(2-methyl-4-oxo-4- phenylbutanoyl)-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC 35% 0.80 min 492 purification (formicacid modifier) (2S,4R)-4-hydroxy-1-(2- methyl-4-oxo-4-phenylbutanoyl)-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during 26% 0.82 min 492 HPLC purification (formicacid modifier) (2S,4R)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1-(2- phenylpropanoyl)pyrrolidine- 2-carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC purification 44% 0.78 min 450 (formicacid modifier) (2S,4R)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1-(2- phenylpropanoyl)pvrrolidine- 2-carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC purification (formic acid modifier) 41%0.80 min 450 (2S,4R)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1-(2-(2-oxopyridin-1(2H)- yl)propanoyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC purification 33% 0.62 min 467 (formicacid modifier) (2S,4R)-4-hydroxy-1-(2- (indolin-1-yl)propanoyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC 35% 0.86 min 491 purification (formicacid modifier) (2S,4R)-4-hydroxy-1-(2- (indolin-1-yl)propanoyl)-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC 40% 0.88 min 491 purification (formicacid modifier) (2S,4R)-4-hydroxy-1-(2- methyl-3-morpholinopropanoyl)-N-(4- (4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

Mixture of diastereo- isomers 57% 0.52 min 472(2S,4R)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1-(pyrazolo[1,5-a]pyridine-2- carbonyl)pyrrolidine-2- carboxamide

64% 0.68 min 463 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- (2-(4-oxoquinazolin-3(4H)-yl)propanoyl)pyrrolidine-2- carboxamide

Mixture of diastereo- isomers 49% 0.71 min 518 (2S,4R)-1-(2-(2,5-dioxopyrrolidin-1- yl)propanoyl)-4-hydroxy-N- (4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC purification 27% 0.60 min 472 (formicacid modifier) (2S,4R)-1-(2-(1H-tetrazol-1-yl)acetyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

73% 0.58 min 428 (2S,4R)-1-(2-(1H-1,2,4- triazol-1-yl)propanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the undefined chiralcentre, eluted first during HPLC 35% 0.58 min 441 purification (formicacid modifier) (2S,4R)-4-hydroxy-1-(2- methyl-3-(1H-1,2,4-triazol-1-yl)propanoyl)-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

Mixture of diastereo- isomers 53% 0.60 min 455(2S,4R)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)-1-((S)-2-(1-oxoisoindolin-2- yl)propanoyl)pyrrolidine-2- carboxamide

Single enantiomer 65% 0.74 min 505 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- ((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2- carboxamide

Single enantiomer 80% 0.76 min 549 (2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1- ((S)-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2- carboxamide

Single enantiomer 57% 0.67 min 519 (2S,4R)-1-((S)-2-cyclopropyl-2-(1-oxoisoindolin-2- yl)acetyl)-4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer 62% 0.80 min 531 (2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(1-oxoisoindolin-2- yl)butanoyl)-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer 58% 0.83 min 533 (2S,4R)-1-((S)-3,3-dimethyl-2-(1-oxoisoindolin-2- yl)butanoyl)-4-hydroxy-N-(4- (4-methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer 52% 0.91 min 547

3-(2-(2-(2-(3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)phenoxy)ethoxy)ethoxy)ethoxy)propanoicacid

An ice-cooled mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (682 mg, 2.2 mmol),3-((14,14-dimethyl-12-oxo-3,6,9,13-tetraoxapentadecyl)oxy)benzoic acid(778 mg, 2.0 mmol), DIPEA (1.36 mL, 7.8 mmol) in DMF (12 mL) was treatedwith HATU (817 mg, 2.2 mmol). The mixture was allowed to warm to ambienttemperature and stirred for 30 minutes then treated with water (70 mL)and extracted with ethyl acetate (3×70 mL). The combined organic phasewas washed with saturated aqueous sodium bicarbonate (100 mL), water(100 mL), brine (100 mL), dried over magnesium sulfate, filtered andevaporated to dryness. The crude product was dissolved indichloromethane (6 mL) and treated with TFA (2.0 mL). After 1 hour, thereaction mixture was evaporated to dryness and the product was purifiedby flash chromatography (60 g C18 cartridge) using a gradient elutionfrom 10 to 95% acetonitrile (+0.1% formic acid) in water (+0.1% formicacid) to afford the title compound (568 mg, 45% yield). LCMS RT=0.73min, ES+ve m/z 642 [M+H]⁺.

(2S,4R)-4-hydroxy-1-(3-(2-methoxyethoxy)benzoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of (2S,4R)-4-hydroxy-1-(3-hydroxybenzoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(55 mg, 0.13 mmol) and potassium carbonate (55 mg, 0.40 mmol) in DMF(0.8 mL) was treated with 1-bromo-2-methoxyethane (commerciallyavailable from for example Aldrich) (0.024 mL, 0.25 mmol) and stirred at50° C. for 2.5 hours. Additional 1-bromo-2-methoxyethane (0.024 mL, 0.25mmol) was added and the mixture stirred at 50° C. overnight. The productwas subjected to purification by mass-directed automated preparativeHPLC (formic acid modifier) to afford the title compound (41 mg, 0.083mmol, 66% yield) LCMS RT=0.74 min, ES+ve m/z 496 [M+H]⁺

(2S,4R)-4-hydroxy-1-(3-(2-(2-methoxyethoxy)ethoxy)benzoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of (2S,4R)-4-hydroxy-1-(3-hydroxybenzoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(55 mg, 0.13 mmol) and potassium carbonate (55 mg, 0.40 mmol) in DMF(0.8 mL) was treated with 1-bromo-2-(2-methoxyethoxy)ethane(commercially available from for example Aldrich) (0.034 mL, 0.25 mmol)and the reaction stirred at 50° C. for 2.5 hours. Additional1-bromo-2-(2-methoxyethoxy)ethane (0.034 mL, 0.25 mmol) was added andthe mixture stirred at 50° C. overnight. The product was subjected topurification by mass-directed automated preparative HPLC (formic acidmodifier) to afford the title compound (40 mg, 0.074 mmol, 59% yield)LCMS RT=0.74 min, ES+ve m/z 540 [M+H]⁺.

(2S,4R)-1-((S)-2-((S)-2-acetamido-4-methylpentanamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-1-((S)-2-aminopropanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (50 mg, 0.12 mmol) and (S)-2-acetamido-4-methylpentanoicacid (commercially available from for example Aldrich) (22 mg, 0.12mmol) in DMF (0.7 mL) was treated with DIPEA (0.062 mL, 0.35 mmol) andthen with HATU (49 mg, 0.13 mmol) and the mixture was stirred at ambienttemperature for 10 minutes. The product was subjected to purification bymass-directed automated preparative HPLC (formic acid modifier) toafford the title compound (42 mg, 0.077 mmol, 66% yield). LCMS RT=0.68min, ES+ve m/z 544 [M+H]⁺.

(2S,4R)-1-((S)-2-acetamido-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A solution of tert-butyl((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)carbamate(120 mg, 0.23 mmol) in dichloromethane (2 mL) and treated with 4 Mhydrochloric acid in 1,4-dioxane (1 mL). The mixture was stirred atambient temperature for 30 minutes and was then evaporated to dryness.The residue was dissolved in DMF (1 mL) and treated with triethylamine(0.08 mL, 0.58 mmol), followed by acetic anhydride (0.02 mL, 0.21 mmol)and the mixture was stirred at ambient temperature for 1 hour. Theproduct was subjected to purification by mass-directed automatedpreparative HPLC (formic acid modifier) to afford the title compound (53mg, 49% yield). LCMS RT=0.65 min, ES+ve m/z 460 [M+H]⁺.

Using a method analogous to that for(2S,4R)-1-((S)-2-acetamido-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,the following compounds were prepared:

Name Structure Yield RT [M + H]+ (2S,4R)-1-((S)-1- acetylpiperidine-2-carbonyl)-4-hydroxy-N- (4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

49% 0.67 min 471 (2S,4R)-1-((S)-4- acetylmorpholine-3-carbonyl)-4-hydroxy-N- (4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2carboxamide

57% 0.59 min 473 (2S,4R)-1-((S)-2- acetamidobutanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

82% 0.61 min 445 (2S,4R)-1-((S)-2- acetamido-3,3- dimethylbutanoyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

68% 0.71 min 473 (2S,4R)-1-((S)-2- acetamido-2- cyclopropylacetyl)-4-hydroxy-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

67% 0.62 min 457

(2S,4R)-1-((S)-2-(3-ethoxy-N-methylbenzamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of (2 S,4R)-4-hydroxy-1-((S)-2-(methylamino)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(20 mg, 0.05 mmol), DIPEA (0.043 mL, 0.25 mmol) and 3-ethoxybenzoic acid(commercially available from for example Aldrich) (8 mg, 0.05 mmol) inDMF (1 mL) was treated with HATU (19 mg, 0.05 mmol) and the mixture wasstirred for 20 minutes. The product was purified by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (14 mg, 0.025 mmol, 50% yield). LCMS RT=0.82 min, ES+ve m/z 551[M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-(3-methoxypropanamido)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A solution of a mixture of(2S,4R)-1-((S)-2-aminopropanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (107 mg, 0.25 mmol), 3-methoxypropionic acid (commerciallyavailable from for example Aldrich) (0.028 mL, 0.30 mmol) and DIPEA (0.2mL, 1.15 mmol) in dry DMF (3 mL) was treated with HATU (115 mg, 0.30mmol). The mixture was stirred at ambient temperature for 30 minutes.The mixture was loaded onto a methanol-preconditioned aminopropylsolid-phase extraction cartridge (2 g), which was eluted with methanol(3 column volumes). The resulting eluant was evaporated to dryness andthe product was subjected to purification by mass-directed automatedpreparative HPLC (formic acid modifier) to afford the title compound (57mg, 0.12 mmol, 48% yield). LCMS RT=0.62 min, ES+ve m/z 475 [M+H]⁺.

(2S,4R)-4-hydroxy-1-(2-(3-methoxypropanamido)-2-methylpropanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A solution of a mixture of(2S,4R)-1-(2-amino-2-methylpropanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(95 mg, 0.24 mmol), 3-methoxypropionic acid (0.028 mL, 0.30 mmol) andDIPEA (0.2 mL, 1.15 mmol) in dry DMF (3 mL) was treated with HATU (115mg, 0.30 mmol) and the mixture was stirred at ambient temperature for 30minutes. The mixture was loaded onto a methanol-preconditionedaminopropyl solid-phase extraction cartridge (NH2) which was eluted withmethanol (3 column volumes). The resulting eluant was evaporated todryness and the product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (45 mg, 0.09 mmol, 37% yield). LCMS RT=0.68 min, ES+ve m/z 489[M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-(2-methoxyacetamido)-3-methylbutanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-1-((S)-2-amino-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (20 mg, 0.044 mmol), 2-methoxyacetic acid (3 μL, 0.039mmol) and DIPEA (0.035 mL, 0.20 mmol) in DMF (1 mL) was treated withHATU (18 mg, 0.047 mmol) and stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (14 mg, 0.029 mmol, 73% yield). LCMS RT=0.70 min, ES+ve m/z 489[M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-(2-methoxy-N-methylacetamido)-3-methylbutanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(methylamino)butanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (19 mg, 0.041 mmol), 2-methoxyacetic acid (3 μL, 0.039mmol) and DIPEA (0.035 mL, 0.20 mmol) in DMF (1 mL) was treated withHATU (18 mg, 0.047 mmol) and stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (16 mg, 0.032 mmol, 81% yield).

LCMS RT=0.70 min, ES+ve m/z 503 [M+H]⁺.

(2S,4R)-1-((S)-2-(N,3-dimethyloxetane-3-carboxamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of (2S,4R)-4-hydroxy-1-((S)-2-(methylamino)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(15 mg, 0.037 mmol), DIPEA (0.032 mL, 0.18 mmol) and3-methyloxetane-3-carboxylic acid (commercially available from forexample Fluorochem) (3 μL, 0.037 mmol) in DMF (1 mL) was treated withHATU (14 mg, 0.037 mmol) and the mixture was stirred for 1 hour. Theproduct was purified by mass-directed automated preparative HPLC (formicacid modifier) to afford the title compound (9 mg, 0.019 mmol, 51%yield). LCMS RT=0.62 min, ES+ve m/z 501 [M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-(3-methyloxetane-3-carboxamido)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-1-((S)-2-aminopropanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(21 mg, 0.043 mmol), DIPEA (0.043 mL, 0.25 mmol) and3-methyloxetane-3-carboxylic acid (commercially available from forexample Fluorochem) (6 mg, 0.05 mmol) in DMF (2 mL) was treated withHATU (19 mg, 0.05 mmol) and stirred for 30 minutes. The product waspurified by mass-directed automated preparative HPLC (formic acidmodifier) to afford the title compound (14 mg, 0.029 mmol, 60% yield).LCMS RT=0.59 min, ES+ve m/z 487 [M+H]⁺.

(2S,4R)-1-((S)-2-(3-ethoxybenzamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of 3-ethoxybenzoic acid (commercially available fromfor example Aldrich) (20 mg, 0.12 mmol) and(2S,4R)-1-((S)-2-aminopropanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (56 mg, 0.13 mmol) in DMF (3.2 mL) was treated with DIPEA(0.063 mL, 0.36 mmol) and then with HATU (50 mg, 0.13 mmol) and themixture was stirred at ambient temperature for 30 minutes. The productwas then subjected to purification by mass-directed automatedpreparative HPLC (formic acid modifier) to afford the title compound (36mg, 0.067 mmol, 56% yield). LCMS RT=0.80 min, ES+ve m/z 537 [M+H]⁺.

(2S,4R)—N-((1H-indol-3-yl)methyl)-4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxamide

A solution of(2S,4R)-4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxylicacid (10 mg, 0.031 mmol), DIPEA (0.038 mL, 0.22 mmol) and(1H-indol-3-yl)methanamine (commercially available from for exampleFluorochem) (6 mg, 0.041 mmol) in DMF (0.8 mL) was treated with HATU (15mg, 0.039 mmol) and stirred for 1 hour. The product was purified bymass-directed automated preparative HPLC (formic acid modifier) toafford the title compound (3.1 mg, 6.9 μmol, 22% yield). LCMS RT=0.75min, ES+ve m/z 447 [M+H]⁺.

(2S,4R)—N—((R)-2,3-dihydrobenzofuran-3-yl)-4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxamide&(2S,4R)—N—((S)-2,3-dihydrobenzofuran-3-yl)-4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxamide

A mixture of 2,3-dihydrobenzofuran-3-amine (commercially available fromfor example Chem-Impex International, Inc.) (13 mg, 0.094 mmol) and(2S,4R)-4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxylicacid (25 mg, 0.079 mmol) in DMF (0.8 mL) was treated with DIPEA (0.055mL, 0.31 mmol) and then HATU (33 mg, 0.086 mmol) and the mixture wasstirred at ambient temperature for 30 minutes. The product mixture wassubjected to purification by mass-directed automated preparative HPLC(formic acid modifier) to afford the title compounds: Isomer 1(first-eluting) (12 mg, 0.027 mmol, 35% yield). LCMS RT=0.73 min, ES+vem/z 436 [M+H]⁺. Isomer 2 (second-eluting) (13 mg, 0.030 mmol, 38%yield). LCMS RT=0.74 min, ES+ve m/z 436 [M+H]⁺.

Using a method analogous to that for the two diastereoisomers of(2S,4R)—N-(2,3-dihydrobenzofuran-3-yl)-4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxamide,the following compounds were prepared:

Name Structure Yield RT [M + H]+ (2S,4R)-N-((S)-1-(4-chlorophenyl)ethyl)-1-(3- ethoxybenzoyl)-4- hydroxypyrrolidine-2-carboxamide

56% 0.92 min 417 (2S,4R)-N-benzyl-1-(3- ethoxybenzoyl)-4-hydroxypyrrolidine-2- carboxamide

64% 0.76 min 369 (2S,4R)-1-(3- ethoxybenzoyl)-4-hydroxy-N-((S)-2-oxopyrrolidin-3- yl)pyrrolidine-2- carboxamide

77% 0.50 min 362 (2S,4R)-1-(3- ethoxybenzoyl)-4-hydroxy-N-((2-methylthiazol-5- yl)methyl)pyrrolidine-2- carboxamide

69% 0.57 min 390 (2S,4R)-1-(3- ethoxybenzoyl)-4-hydroxy-N-((1-methyl-1H-pyrazol-4- yl)methyl)pyrrolidine-2- carboxamide

70% 0.55 min 375 (2S,4R)-N-([1,1′-biphenyl]- 4-ylmethyl)-1-(3-ethoxybenzoyl)-4- hydroxypyrrolidine-2- carboxamide

55% 0.98 min 445 (2S,4R)-1-(3- ethoxybenzoyl)-4-hydroxy-N-((5-phenyl-1,2,4- oxadiazol-3- yl)methyl)pyrrolidine-2- carboxamide

60% 0.82 min 437 (2S,4R)-N-((3-(4- chlorophenyl)-1,2,4-oxadiazol-5-yl)methyl)-1- (3-ethoxybenzoyl)-4- hydroxypyrrolidine-2-carboxamide

47% 0.95 min 471 (2S,4R)-1-(3- ethoxybenzoyl)-4-hydroxy-N-((5-phenylisoxazol-3- yl)methyl)pyrrolidine-2- carboxamide

60% 0.86 min 436 (2S,4R)-1-(3- ethoxybenzoyl)-4-hydroxy-N-((1-methyl-1,2,3,4- tetrahydroquinolin-6- yl)methyl)pyrrolidine-2-carboxamide

30% 0.67 min 438

(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (125 mg, 0.34 mmol) and(S)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoic acid (83 mg, 0.36 mmol) inDMF (1.6 mL) was treated with DIPEA (0.24 mL, 1.4 mmol) and HATU (140mg, 0.37 mmol) and the mixture was stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (120 mg, 0.22 mmol, 65% yield). LCMS RT=0.81 min, ES+ve m/z 549[M+H]⁺.

(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)-1-((R)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (65 mg, 0.18 mmol) and(R)-3-methyl-2-(1-oxoisoindolin-2-yl)butanoic acid (43 mg, 0.19 mmol) inDMF (1.6 mL) was treated with DIPEA (0.123 mL, 0.70 mmol) and HATU (74mg, 0.19 mmol) and the mixture was stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (64 mg, 0.12 mmol, 66% yield). LCMS RT=0.80 min, ES+ve m/z 549[M+H]⁺.

tert-butyl((S)-1-((2R,4S)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-yl)(methyl)carbamate

A stirred mixture of (S)-2-((tert-butoxycarbonyl)(methyl)amino)propanoicacid (115 mg, 0.57 mmol) and(2R,4S)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (200 mg, 0.57 mmol) in DMF (0.7 mL) was treated with DIPEA(0.4 mL, 2.3 mmol) and then with HATU (215 mg, 0.57 mmol) and themixture was stirred at ambient temperature for 30 minutes. The productwas subjected to purification by mass-directed automated preparativeHPLC (formic acid modifier) to afford the title compound (146 mg, 0.29mmol, 51% yield). LCMS RT=0.84 min, ES+ve m/z 503 [M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-(3-methoxy-N-methylpropanamido)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-4-hydroxy-1-((S)-2-(methylamino)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(50 mg, 0.12 mmol) and 3-methoxypropanoic acid (commercially availablefrom for example Aldrich) (0.013 mL, 0.14 mmol) in DMF (0.8 mL) wastreated with DIPEA (0.087 mL, 0.50 mmol) and then with HATU (52 mg, 0.14mmol). The mixture was stirred at ambient temperature for 30 minutes.The product was subjected to purification by mass-directed automatedpreparative HPLC (formic acid modifier) to afford the title compound (43mg, 71% yield). LCMS RT=0.61 min, ES+ve m/z 489 [M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-((2-methoxyethyl)(methyl)amino)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of 1-bromo-2-methoxyethane (commercially availablefrom for example Aldrich) (0.013 mL, 0.14 mmol) and(2S,4R)-4-hydroxy-1-((S)-2-(methylamino)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(50 mg, 0.12 mmol) in DMF (0.8 mL) was treated with DIPEA (0.054 mL,0.31 mmol) and the mixture was stirred at 85° C. for 18 hours. Thereaction mixture was cooled and the product was subj ected topurification by mass-directed automated preparative HPLC (ammoniumbicarbonate modifier) to afford the title compound (44 mg, 77% yield).LCMS RT=0.51 min, ES+ve m/z 461 [M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-4-(2-methoxyacetyl)morpholine-3-carbonyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1-((S)-morpholine-3-carbonyl)pyrrolidine-2-carboxamide,hydrochloride (19 mg, 0.041 mmol), 2-methoxyacetic acid (3 μL, 0.039mmol) and DIPEA (0.035 mL, 0.20 mmol) in DMF (1 mL) was treated withHATU (18 mg, 0.047 mmol) and stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (13 mg, 0.026 mmol, 66% yield). LCMS RT=0.60 min, ES+ve m/z 503[M+H]⁺.

(2S,4R)—N-(4-(2,4-dimethylthiazol-5-yl)benzyl)-4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)—N-(4-(2,4-dimethylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide(30 mg, 0.09 mmol) and 2-(3-methylisoxazol-5-yl)acetic acid(commercially available from for example Aldrich) (13 mg, 0.09 mmol) inDMF (0.8 mL) was treated with DIPEA (0.063 mL, 0.36 mmol) and then withHATU (41 mg, 0.11 mmol) and stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (26 mg, 63% yield). LCMS RT=0.66 min, ES+ve m/z 455 [M+H]⁺.

(2S,4R)-1-((S)-2-acetamidopropanoyl)-N-(4-(2,4-dimethylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)—N-(4-(2,4-dimethylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide(30 mg, 0.09 mmol) and (S)-2-acetamidopropanoic acid (commerciallyavailable from for example Aldrich) (12 mg, 0.09 mmol) in DMF (0.8 mL)was treated with DIPEA (0.063 mL, 0.36 mmol) and then with HATU (41 mg,0.11 mmol), and the mixture was stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (30 mg, 75% yield). LCMS RT=0.58 min, ES+ve m/z 445 [M+H]⁺.

(2S,4R)—N-(4-bromobenzyl)-1-(3-ethoxybenzoyl)-4-hydroxypyrrolidine-2-carboxamide

An ice-cooled mixture of(2S,4R)-1-(3-ethoxybenzoyl)-4-hydroxypyrrolidine-2-carboxylic acid (73mg, 0.26 mmol) and (4-bromophenyl)methanamine, hydrochloride(commercially available from for example Aldrich) (58 mg, 0.26 mmol) inDMF (0.5 mL) was treated with a solution of DIPEA (0.145 mL, 0.83 mmol)in DMF (1 mL) and then with HATU (105 mg, 0.28 mmol) and stirredovernight at ambient temperature. The product was then subjected topurification by mass-directed automated preparative HPLC (formic acidmodifier) to afford the title compound (62 mg, 53% yield). LCMS RT=0.90min, ES+ve m/z 447,449 [M+H]⁺.

(2S,4R)—N-([1,1′-biphenyl]-4-ylmethyl)-1-(3-ethoxybenzoyl)-4-hydroxypyrrolidine-2-carboxamide

A mixture of (2S,4R)-1-(3-ethoxybenzoyl)-4-hydroxypyrrolidine-2-carboxylic acid (30 mg,0.11 mmol) and [1,1′-biphenyl]-4-ylmethanamine (commercially availablefrom for example Aldrich) (20 mg, 0.11 mmol) in DMF (0.8 mL) was treatedwith DIPEA (0.075 mL, 0.43 mmol) and then with HATU (45 mg, 0.12 mmol)and stirred at ambient temperature for 30 minutes. The product wassubjected to purification by mass-directed automated preparative HPLC(formic acid modifier) to afford the title compound (26 mg, 55% yield).LCMS RT=0.98 min, ES+ve m/z 445 [M+H]⁺.

Using a method analogous to that for2S,4R)—N-([1,1′-biphenyl]-4-ylmethyl)-1-(3-ethoxybenzoyl)-4-hydroxypyrrolidine-2-carboxamide,the following compounds were prepared:

Name Structure Yield RT [M + H]+ (2S,4R)-N-((S)-1-(4-chlorophenyl)ethyl)-1-(3- ethoxybenzoyl)-4- hydroxypyrrolidine-2-carboxamide

56% 0.92 min 417 (2S,4R)-N-benzyl-1-(3- ethoxybenzoyl)-4-hydroxypyrrolidine-2- carboxamide

64% 0.76 min 369 (2S,4R)-1-(3- ethoxybenzoyl)-4- hydroxy-N-((S)-2-oxopyrrolidin-3- yl)pyrrolidine-2- carboxamide

77% 0.50 min 362 (2S,4R)-1-(3- ethoxybenzoyl)-4- hydroxy-N-((2-methylthiazol-5- yl)methyl)pyrrolidine-2- carboxamide

69% 0.57 min 390 (2S,4R)-1-(3- ethoxybenzoyl)-4- hydroxy-N-((1-methyl-1H-pyrazol-4- yl)methyl)pyrrolidine-2- carboxamide

70% 0.55 min 375 (2S,4R)-N-([1,1′- biphenyl]-4-ylmethyl)-1-(3-ethoxybenzoyl)-4- hydroxypyrrolidine-2- carboxamide

55% 0.98 min 445 (2S,4R)-1-(3- ethoxybenzoyl)-4- hydroxy-N-((5-phenyl-1,2,4-oxadiazol-3- yl)methyl)pyrrolidine-2- carboxamide

60% 0.82 min 437 (2S,4R)-N-((3-(4- chlorophenyl)-1,2,4-oxadiazol-5-yl)methyl)-1- (3-ethoxybenzoyl)-4- hydroxypyrrolidine-2-carboxamide

47% 0.95 min 471 (2S,4R)-1-(3- ethoxybenzoyl)-4- hydroxy-N-((5-phenylisoxazol-3- yl)methyl)pyrrolidine-2- carboxamide

60% 0.86 min 436 (2S,4R)-1-(3- ethoxybenzoyl)-4- hydroxy-N-((1-methyl-1,2,3,4- tetrahydroquinolin-6- yl)methyl)pyrrolidine-2- carboxamide

30% 0.67 min 438

(2S,4R)-4-hydroxy-1-(2-(3-methoxypropanamido)acetyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-1-(2-aminoacetyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(134 mg, 0.33 mmol) and 3-methoxypropanoic acid (commercially availablefrom for example Aldrich) (37 mg, 0.36 mmol) in DMF (0.8 mL) was treatedwith DIPEA (0.23 mL, 1.3 mmol) and then with HATU (136 mg, 0.36 mmol)and stirred at ambient temperature for 30 minutes. The product wassubjected to purification by mass-directed automated preparative HPLC(formic acid modifier) to afford the title compound (36 mg, 24% yield).LCMS RT=0.56 min, ES+ve m/z 461 [M+H]⁺.

(2S,4R)-1-((S)-3,3-dimethyl-2-(3-methyloxetane-3-carboxamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (20 mg, 0.04 mmol) and 3-methyloxetane-3-carboxylic acid(commercially available from for example Chemgenx) (5 mg, 0.04 mmol) inDMF (0.6 mL) was treated with DIPEA (0.03 mL, 0.17 mmol) and then withHATU (20 mg, 0.05 mmol), and the mixture was stirred at ambienttemperature for 1 hour. The product was subjected to purification bymass-directed automated preparative HPLC (formic acid modifier) to givethe title compound (18 mg, 80% yield). LCMS RT=0.76 min, ES+ve m/z 529[M+H]⁺.

Using a method analogous to that for(2S,4R)-1-((S)-3,3-dimethyl-2-(3-methyloxetane-3-carboxamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,the following compounds were prepared:

Name Structure Yield RT [M + H]+ (2S,4R)-1-((S)-3,3-dimethyl-2-(oxetane-3- carboxamido)butanoyl)-4- hydroxy-N-(4-(4-methylthiazol-5- yl)benzyl)pyrrolidine-2- carboxamide

41% 0.72 min 515 (2S,4R)-1-((S)-2- (cyclopentanecarboxamido)-3,3-dimethylbutanoyl)- 4-hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

58% 0.89 min 527 (2S,4R)-1-((S)-3,3- dimethyl-2-(tetrahydro- 2H-pyran-4-carboxamido)butanoyl)-4- hydroxy-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

52% 0.76 min 543

(S)—N—((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)morpholine-3-carboxamide,hydrochloride

A mixture of (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(40 mg, 0.086 mmol) and(S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (commerciallyavailable from for example Astatech, Inc.) (20 mg, 0.086 mmol) in DMF(0.6 mL) was treated with DIPEA (0.06 mL, 0.35 mmol) and then with HATU(40 mg, 0.10 mmol) and stirred at ambient temperature for 1 hour. Theproduct was subjected to purification by mass-directed automatedpreparative HPLC (formic acid modifier) to give the intermediateBoc-protected product. The intermediate was then dissolved in a mixtureof dichloromethane (1 mL) and methanol (0.5 mL) and treated with 4Mhydrochloric acid in 1,4-dioxane (0.4 mL, 1.6 mmol), After stirring atambient temperature for 1 hour, the mixture was evaporated to dryness toafford the title compound (31 mg, 62% yield). LCMS RT=0.60 min, ES+vem/z 544 [M+H]⁺.

Using a method analogous to that for(S)—N—((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)morpholine-3-carboxamide,hydrochloride, the following compounds were prepared:

(R)-N-((S)-1-((2S,4R)-4- hydroxy-2-((4-(4- methylthiazol-5-yl)benzyl)carbamoyl) pyrrolidin- 1-yl)-3,3-dimethyl-1-oxobutan-2-yl)morpholine-3- carboxamide, hydrochloride

58% 0.63 min 544 (S)-N-((S)-1-((2S,4R)-4- hydroxy-2-((4-(4-methylthiazol-5- yl)benzyl)carbamoyl) pyrrolidin- 1-yl)-3,3-dimethyl-1-oxobutan-2-yl)morpholine-2- carboxamide, hydrochloride

54% 0.62 min 544 N-((S)-1-((2S,4R)-4-hydroxy- 2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl) pyrrolidin- 1-yl)-3,3-dimethyl-1-oxobutan-2-yl)morpholine-2- carboxamide, hydrochloride

52%  0.62- 0.64 min 544

tert-butyl4-((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-yl)piperazine-1-carboxylate& tert-butyl4-((R)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-yl)piperazine-1-carboxylate

A stirred mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (100 mg, 0.28 mmol) and2-(4-(tert-butoxycarbonyl)-2-oxopiperazin-1-yl)propanoic acid (85 mg,0.31 mmol) in DMF (0.8 mL) was treated with DIPEA (0.20 mL, 1.13 mmol)and then with HATU (129 mg, 0.34 mmol) and then stirred at ambienttemperature for 30 minutes. The product was subjected to purification bymass-directed automated preparative HPLC (ammonium bicarbonate modifier)to afford the title compounds: Isomer 1 (first-eluting) (48 mg, 30%yield). LCMS RT=0.85 min, ES+ve m/z 572 [M+H]⁺. Isomer 2(second-eluting) (51 mg, 32% yield). LCMS RT=0.86 min, ES+ve m/z 572[M+H]⁺.

(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(piperazin-1-yl)propanoyl)pyrrolidine-2-carboxamide,hydrochloride &(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1-((R)-2-(piperazin-1-yl)propanoyl)pyrrolidine-2-carboxamide,hydrochloride

Isomer 1 and isomer 2 of tert-butyl4-(1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-yl)piperazine-1-carboxylate(48 mg, 0.08 mmol) were separately dissolved in a mixture ofdichloromethane (0.3 mL) and methanol (0.1 mL) and treated with 4Mhydrochloric acid in 1,4-dioxane (0.3 mL, 1.2 mmol) respectively. Afterstirring at ambient temperature for 1 hour, the reaction mixtures wereevaporated to dryness to afford the title compounds as hydrochloridesalts. Isomer 1 (42 mg, 99% yield). LCMS RT=0.62 min, ES+ve m/z 472[M+H]⁺. Isomer 2 (42 mg, 99% yield). LCMS RT=0.60 min, ES+ve m/z 472[M+H]⁺.

(S)—N—((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)-4-(2-methoxyethyl)morpholine-2-carboxamide

A mixture of 1-bromo-2-methoxyethane (4 μL, 0.04 mmol),(S)—N—((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)morpholine-2-carboxamide,hydrochloride (20 mg, 0.04 mmol) and DIPEA (0.019 mL, 0.11 mmol) in DMF(0.5 mL) was stirred at 85° C. for 6 hours. The cooled product wassubjected to purification by mass-directed automated preparative HPLC(ammonium bicarbonate modifier) to afford the title compound (9 mg, 41%yield). LCMS RT=0.88 min, ES+ve m/z 602 [M+H]⁺.

Using a method analogous to that for(S)—N—((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)-4-(2-methoxyethyl)morpholine-2-carboxamidethe following compounds were prepared:

Stereo- chemistry [M + Name Structure Comments Yield RT H]+(2S,4R)-4-hydroxy-1-(2-(4- (2-methoxyethyl)-2- oxopiperazin-1-yl)propanoyl)-N-(4-(4- methylthiazol-5- yl)benzyl)pyrrolidine-2-carboxamide

Single enantiomer, stereo- chemistry unknown at the unspecified chiralcentre 20% 0.68 min 530 (2S,4R)-4-hydroxy-1-(2-(4- (2-methoxyethyl)-2-oxopiperazin-1- yl)propanovl)-N-(4-(4- methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide

Single enantiomer, stereo- chemistry unknown at the unspecified chiralcentre 32% 0.69 min 530 (S)-N-((S)-1-((2S,4R)-4- hydroxy-2-((4-(4-methylthiazol-5- yl)benzyl)carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)-4-(2- methoxyethyl)morpholine- 3-carboxamide

41% 0.86 min 602 (R)-N-((S)-1-((2S,4R)-4- hydroxy-2-((4-(4-methylthiazol-5- yl)benzyl)carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)-4-(2- methoxyethyl)morpholine- 3-carboxamide

45% 0.86 min 602

Methyl4-(((S)-1-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)amino)-4-oxobutanoate

A mixture of(2S,4R)-1-((S)-2-((S)-2-amino-4-methylpentanamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (102 mg, 0.19 mmol), DIPEA (0.165 mL, 0.95 mmol) and4-methoxy-4-oxobutanoic acid (commercially available from for exampleAldrich) (25 mg, 0.19 mmol) in DMF (2 mL) was treated with HATU (64 mg,0.17 mmol) and the mixture was stirred at ambient temperature for 20minutes. Brine (10 mL) was added and the product was extracted withethyl acetate (20 mL). The organic phase was washed with brine (2×20mL), dried using a hydrophobic frit and evaporated to dryness. Theproduct was purified by chromatography on reverse phase silica using agradient elution from 5% to 70% acetonitrile (+0.1% formic acid) inwater (+0.1% formic acid) to afford the title compound (73 mg, 0.12mmol, 63% yield). LCMS RT=0.74 min, ES+ve m/z 616 [M+H]⁺.

4-(3-((2S,4R)-4-Hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)phenoxy)butanoicacid (

A solution of tert-butyl4-(3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)phenoxy)butanoate(130 mg, 0.22 mmol) in dichloromethane (3 mL) was treated with TFA (0.5mL, 6.5 mmol) and stirred at ambient temperature for 5 hours. Thesolvent was evaporated to dryness and the product was subjected topurification by mass-directed automated preparative HPLC (formic acidmodifier) to afford the title compound (65 mg, 0.12 mmol, 55% yield).LCMS RT=0.70 min, ES+ve m/z 524 [M+H]⁺.

4-(((S)-1-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)amino)-4-oxobutanoicacid

A solution of methyl4-(((S)-1-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-1-oxopropan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)amino)-4-oxobutanoate(73 mg, 0.12 mmol) in methanol (3 mL) was treated with aqueous sodiumhydroxide (2M, 0.6 mL, 1.2 mmol) and the mixture was stirred at ambienttemperature for 2 hours. The mixture was evaporated to dryness and theproduct was purified by chromatography on reverse phase silica using agradient elution from 5% to 60% acetonitrile (+0.1% formic acid) inwater (+0.1% formic acid) to afford the title compound (53 mg, 0.088mmol, 74% yield). LCMS RT=0.69 min, ES+ve m/z 602 [M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-((S)-2-(2-methoxyacetamido)-4-methylpentanamido)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-1-((S)-2-((S)-2-amino-4-methylpentanamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (30 mg, 0.056 mmol), 2-methoxyacetic acid (commerciallyavailable from for example Aldrich) (4.3 uL, 0.056 mmol) and DIPEA (0.05mL, 0.29 mmol) in DMF (1 mL) was treated with HATU (25 mg, 0.066 mmol)and stirred at ambient temperature for 30 minutes. The product wassubjected to purification by mass-directed automated preparative HPLC(formic acid modifier) to afford the title compound (18 mg, 0.031 mmol,56% yield). LCMS RT=0.73 min, ES+ve m/z 574 [M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-((S)-2-(3-methoxypropanamido)-4-methylpentanamido)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxaide

A mixture of(2S,4R)-1-((S)-2-((S)-2-amino-4-methylpentanamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (30 mg, 0.056 mmol), 3-methoxypropanoic acid (commerciallyavailable from for example Aldrich) (5.2 uL, 0.056 mmol) and DIPEA (0.05mL, 0.29 mmol) in DMF (1 mL) and stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (20 mg, 0.034 mmol, 61% yield). LCMS RT=0.72 min, ES+ve m/z 588[M+H]⁺.

(2S,4R)-1-(6-cyanopyridin-2-yl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (58 mg, 0.16 mmol) and 6-fluoropicolinonitrile(commercially available from for example Aldrich) (20 mg, 0.16 mmol) inDMSO (1 mL) was treated with DIPEA (0.10 mL, 0.57 mmol), sealed andheated in a Biotage “Initiator” microwave at 100° C. for 60 minutes. Theproduct was purified by mass-directed automated preparative HPLC (formicacid modifier) to afford the title compound (23 mg, 0.055 mmol, 34%yield). LCMS RT=0.74 min, ES+ve m/z 420 [M+H]⁺.

Intermediates 4-(oxazol-5-yl)benzonitrile

A mixture of 4-formylbenzonitrile (commercially available from forexample Aldrich) (5.32 g, 41 mmol),1-((isocyanomethyl)sulfonyl)-4-methylbenzene (commercially availablefrom for example Aldrich) (8.83 g, 45 mmol) and potassium carbonate (7.3g, 53 mmol) in methanol (200 mL) was stirred at ambient temperature for80 minutes. The mixture was then evaporated to dryness; the residue wastreated with saturated aqueous sodium bicarbonate (100 mL) and extractedwith dichloromethane (3×100 mL). The combined organics were washed withbrine (75 mL), passed through a hydrophobic frit and then evaporated todryness to afford the title compound (7.19 g, 42 mmol, quantitative).LCMS RT=0.48 min, ES+ve m/z 171 [M+H]⁺.

(4-(oxazol-5-yl)phenyl)methanamine

Under an atmosphere of nitrogen, an ice-cooled mixture of4-(oxazol-5-yl)benzonitrile (900 mg, 5.29 mmol) and cobalt(II) chloridehexahydrate (commercially available from for example Aldrich) (1.8 g,7.9 mmol) in methanol (50 mL) was treated portion-wise over 5 minuteswith sodium borohydride (1 g, 26 mmol). The mixture was stirred for 30minutes and then treated with water (50 mL) and concentrated aqueousammonia (20 mL). The mixture was extracted with chloroform (3×150 mL),the combined organics were evaporated to dryness and the product waspurified by chromatography on silica using a gradient elution from 0% to30% methanol in dichloromethane (+0.1% triethylamine) to afford thetitle compound (580 mg, 3.3 mmol, 63% yield). LCMS RT=0.35 min, ES+vem/z 175 [M+H]⁺.

(2S,4R)-tert-butyl4-hydroxy-2-((4-(oxazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate

To a stirred solution of(2S,4R)-1-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid(0.66 g, 2.9 mmol) in dry DMF (20 mL) were added(4-(oxazol-5-yl)phenyl)methanamine (0.5 g, 2.87 mmol) and DIPEA (1 mL,5.7 mmol). This solution was then ice-cooled and HATU (1.09 g, 2.9 mmol)was added. The reaction mixture was stirred with cooling for anadditional hour then treated with water (30 mL) and extracted with ethylacetate (3×100 mL). The combined organic phase was washed with saturatedaqueous sodium bicarbonate (60 mL), brine (60 mL), dried over magnesiumsulfate, filtered and evaporated to dryness. The product was purified bychromatography on silica using a gradient elution from 0% to 25%methanol in dichloromethane to afford the title compound (758 mg, 1.96mmol, 68% yield). LCMS RT=0.73 min, ES+ve m/z 388 [M+H]⁺.

(2S,4R)-4-hydroxy-N-(4-(oxazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride

A solution of (2S,4R)-tert-butyl4-hydroxy-2-((4-(oxazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate(2.74 g, 7.1 mmol) in methanol (10 mL) and dichloromethane (15 mL) wastreated with hydrochloric acid (4 M in 1,4-dioxane) (8.8 mL, 35 mmol)and the mixture was stirred at ambient temperature for 24 hours. Themixture was evaporated to dryness. The residue was suspended inmethanol, filtered and dried under vacuum to afford the title compound(2.24 g, 6.9 mmol, 98% yield). LCMS RT=0.44 min, ES+ve m/z 288 [M+H]⁺.

(2S,4R)-tert-butyl2-((4-bromobenzyl)carbamoyl)-4-hydroxypyrrolidine-1-carboxylate

An ice-cooled mixture of(2S,4R)-1-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid(commercially available from for example Aldrich) (7.95 g, 34 mmol) and(4-bromophenyl)methanamine (commercially available from for exampleFluorochem) (6.4 g, 34 mmol) in DMF (200 mL) was treated with DIPEA (18mL, 103 mmol) and then with HATU (14.4 g, 38 mmol) and the mixture wasstirred at ambient temperature for 30 minutes. The mixture was treatedwith water (200 mL) and extracted with ethyl acetate (2×200 mL). Thecombined organic phase was washed with saturated aqueous sodiumbicarbonate (2×300 mL), water (100 mL), brine (200 mL), dried overmagnesium sulfate and evaporated to dryness. The product was purified byflash chromatography (750 g silica cartridge) using a gradient elutionfrom 0% to 10% methanol in dichloromethane to afford the title compound(12.9 g, 94% yield). LCMS RT=0.87 min, ES+ve m/z 401 [M+H]⁺.

(2S,4R)-tert-butyl4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate

Under an atmosphere of nitrogen, a mixture of (2S,4R)-tert-butyl2-((4-bromobenzyl)carbamoyl)-4-hydroxypyrrolidine-1-carboxylate (12.9 g,32 mmol), 4-methylthiazole (commercially available from for exampleAldrich) (5.9 mL, 65 mmol), palladium(II) acetate (commerciallyavailable from for example Aldrich) (0.145 g, 0.65 mmol) and potassiumacetate (6.34 g, 65 mmol) in N-methyl-2-pyrrolidone (80 mL) was stirredat 120° C. for 18 hours. After cooling to ambient temperature, water(100 ml) was added and the product was extracted with ethyl acetate(4×300 mL). The combined organic phase was washed with brine (5×200 mL),dried over magnesium sulfate and evaporated to dryness. The product waspurified by flash chromatography (750 g silica cartridge) using agradient elution from 0% to 10% methanol in dichloromethane to affordthe title compound (8.0 g, 59% yield). LCMS RT=0.75 min, ES+ve m/z 418[M+H]⁺.

(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride

A solution of (2S,4R)-tert-butyl4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate(8 g, 19 mmol) in a mixture of methanol (30 mL) and dichloromethane (20mL) was treated with 4M hydrochloric acid in 1,4-dioxane (8 mL, 32mmol). The mixture was stirred at ambient temperature for 2 hours. Thesolvent was evaporated to dryness and the residue was triturated indichloromethane, filtered and dried under vacuum to afford the titlecompound (6.7 g, 99% yield). LCMS RT=0.51 min, ES+ve m/z 318 [M+H]⁺.

(2S,4R)-1-(3-ethoxybenzoyl)-4-hydroxypyrrolidine-2-carboxylic acid

3-Ethoxybenzoic acid (commercially available from for example Aldrich)(4 g, 24 mmol) was dissolved in thionyl chloride (24 mL, 329 mmol) andstirred at 60° C. for 1 hour and then at 50° C. for 18 hours. Aftercooling to ambient temperature the mixture was evaporated to dryness andthe residue was treated with diethyl ether (5 mL). The mixture was thenice-cooled and treated with a solution of(2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid, hydrochloride(commercially available from for example Aldrich) (4.44 g, 27 mmol) in1M aqueous sodium hydroxide (27 mL, 27 mmol). The reaction was warmed toambient temperature and stirred for 18 hours. The mixture was separated;the aqueous phase was washed with diethyl ether and then acidified with2M aqueous hydrochloric acid. The product was extracted in diethyl ether(2×70 mL) and the combined ethereal phase was evaporated to dryness. Theproduct was purified by flash chromatography (340 g C18 cartridge),using a gradient elution from 10 to 30% acetonitrile (+0.1% formic acid)in water (+0.1% formic acid) to afford the title compound (3.5 g, 52%yield). LCMS RT=0.55 min, ES+ve m/z 280 [M+H]⁺.

(2S,4R)-benzyl1-((S)-2-acetamidopropanoyl)-4-hydroxypyrrolidine-2-carboxylate

An ice-cooled mixture of (S)-2-acetamidopropanoic acid (commerciallyavailable from for example Aldrich) (2.80 g, 21 mmol) and (2S,4R)-benzyl4-hydroxypyrrolidine-2-carboxylate, hydrochloride (commerciallyavailable from for example Aldrich) (5 g, 19 mmol) in DMF (5 mL) wastreated with DIPEA (14 mL, 78 mmol), followed by HATU (8.11 g, 21 mmol)over 10 min. The mixture was warmed to ambient temperature and stirredfor 1 hour then treated with saturated aqueous sodium bicarbonate (30mL) and stirred for 5 min. The mixture was then extracted with ethylacetate (3×100 mL) and the combined organic phase was washed with water(100 mL), brine (100 mL), dried over magnesium sulfate and evaporated todryness. The product was purified by flash chromatography (330 g silicacartridge) using a gradient elution from 0 to 10% methanol indichloromethane to afford the title compound (2.0 g, 31% yield). LCMSRT=0.63 min, ES+ve m/z 335 [M+H]⁺.

(2S,4R)-1-((S)-2-acetamidopropanoyl)-4-hydroxypyrrolidine-2-carboxylicacid

A solution of(2S,4R)-benzyl-1-((S)-2-acetamidopropanoyl)-4-hydroxypyrrolidine-2-carboxylate(2 g, 6.0 mmol) in ethanol (10 mL) was added to a flask containingpalladium on carbon (1.27 g, 1.2 mmol) (10%, Degussa type) under anatmosphere of nitrogen. The flask was filled with hydrogen and thesolution was stirred at ambient temperature for 2 hours. The catalystwas removed by filtration through celite and the filtrate was evaporatedunder reduced pressure to afford the title compound (1.37 g, 94% yield).LCMS RT=0.28 min, ES+ve m/z 244 [M+H]⁺.

(2S,4R)-4-hydroxy-1-(2-3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylicacid

Under an atmosphere of nitrogen, a solution of (2S,4R)-benzyl4-hydroxy-1-(2-(3-methylisoxazol-5-yl)acetyl)pyrrolidine-2-carboxylate(2.3 g, 6.7 mmol) in ethanol (60 mL) was added to palladium on carbon(0.071 g, 0.67 mmol) (10%, Degussa type) and then stirred under anatmosphere of hydrogen. After 2 hours, the mixture was filtered throughcelite. The filtrate was evaporated to dryness and the residue wastriturated with cyclohexane and dried under vacuum to afford a whitesolid. The product was purified by mass-directed automated preparativeHPLC (TFA modifier) to afford the title compound (650 mg, 2.6 mmol, 38%yield). LCMS RT=0.38 min, ES+ve m/z 255 [M+H]⁺.

(2S,4R)-methyl4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxylate

A mixture of (2S,4R)-methyl 4-hydroxypyrrolidine-2-carboxylate,hydrochloride (commercially available from for example Aldrich) (1.77 g,9.8 mmol) and (S)-2-(1-oxoisoindolin-2-yl)propanoic acid (2 g, 9.8 mmol)in DMF (4 mL) was treated with DIPEA (5.11 mL, 29 mmol) and then withHATU (4.08 g, 10.7 mmol), and stirred at ambient temperature for 30minutes. The mixture was treated with saturated aqueous sodiumbicarbonate (100 mL) and extracted with ethyl acetate (2×200 mL). Thecombined organic phase was washed with water (100 mL), brine (100 mL),dried over magnesium sulfate and evaporated to dryness. The product waspurified by flash chromatography (120 g C18 cartridge), using a gradientelution from 10% to 50% acetonitrile (+0.1% formic acid) in water (+0.1%formic acid) to afford the title compound (1.0 g, 31% yield). LCMSRT=0.60 min, ES+ve m/z 333 [M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxylicacid

A solution of (2S,4R)-methyl4-hydroxy-1-((S)-2-(1-oxoisoindolin-2-yl)propanoyl)pyrrolidine-2-carboxylate(1 g, 3.0 mmol) in methanol (2 mL) was treated with 2M aqueous sodiumhydroxide (5 mL, 10 mmol) and the mixture was stirred at ambienttemperature for 2 hours then acidified with 2M aqueous hydrochloric acid(6 mL). The mixture was then evaporated to about one half of theoriginal volume and then ice-cooled. The resulting precipitate wasfiltered off and dried under vacuum to afford the title compound (615mg, 64% yield). LCMS RT=0.51 min, ES+ve m/z 319 [M+H]⁺.

Methyl 3-(4-(tert-butoxy)-4-oxobutoxy)benzoate

A solution of methyl 3-hydroxybenzoate (commercially available from forexample Aldrich) (1 g, 6.6 mmol) and K₂CO₃ (1.82 g, 13.2 mmol) in DMF(10 mL) was treated with tert-butyl 4-bromobutanoate (commerciallyavailable from for example Aldrich) (2.2 g, 9.9 mmol) and the mixturewas stirred at 60° C. for 16 hours. A further aliquot of K₂CO₃ (1.82 g,13.2 mmol) and tert-butyl 4-bromobutanoate (2.2 g, 9.9 mmol) were addedand the mixture was heated at 60° C. for further 6 hours. The mixturewas cooled to ambient temperature and partitioned between ethyl acetate(50 mL) and water (50 mL). The organic phase was washed with brine (2×50mL), dried (hydrophobic frit) and evaporated to dryness. The product waspurified by chromatography on silica using a gradient elution from 0% to100% methyl tert-butyl ether in cyclohexane to afford the title compound(1.4 g, 4.8 mmol, 72% yield). LCMS RT=1.26 min, ES+ve m/z 312 [M+H]⁺.

3-(4-(Tert-butoxy)-4-oxobutoxy)benzoic acid

A mixture of methyl 3-(4-(tert-butoxy)-4-oxobutoxy)benzoate (1.4 g, 4.8mmol) and aqueous sodium hydroxide (2M, 4.8 mL, 9.6 mmol) in methanol(10 mL) was stirred at ambient temperature for 5 hours. The methanol wasremoved under reduced pressure (no heat) and the aqueous phase wasacidified to pH 3 with saturated aqueous citric acid. The product wasextracted with ethyl acetate (60 mL) and the organic extract was washedwith brine (20 mL), dried using a hydrophobic frit and evaporated todryness. The product was purified by chromatography on silica using agradient elution from 0% to 25% methanol in dichloromethane to affordthe title compound (625 mg, 2.2 mmol, 47% yield). LCMS RT=1.06 min,ES+ve m/z 279 [M−H]⁺.

Methyl3-((14,14-dimethyl-12-oxo-3,6,9,13-tetraoxapentadecyl)oxy)benzoate

An ice-cooled mixture of tert-butyl3-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)propanoate (commerciallyavailable from for example Aldrich) (2.0 g, 7.2 mmol),triphenylphosphine (2.3 g, 8.6 mmol) and methyl 3-hydroxybenzoate(commercially available from for example Aldrich) (1.2 g, 7.9 mmol) inTHF (40 mL) was treated dropwise over 5 minutes with diisopropylazodicarboxylate (1.68 mL, 8.6 mmol). The mixture was warmed to ambienttemperature and stirred for 18 hours. The mixture was then evaporated todryness and purified by flash column chromatography (100 g silicacartridge) using a gradient elution from 0 to 100% methyl tert-butylether in cyclohexane over 40 minutes to afford the title compound (2.53g, 85% yield). LCMS RT=1.14 min, ES+ve m/z 430 [M+NH₄]⁺.

3-((14,14-dimethyl-12-oxo-3,6,9,13-tetraoxapentadecyl)oxy)benzoic acid (

A solution of methyl3-((14,14-dimethyl-12-oxo-3,6,9,13-tetraoxapentadecyl)oxy)benzoate (2.53g, 4.9 mmol) in methanol (25 mL) was treated with 1M aqueous sodiumhydroxide (0.3 g, 7.6 mmol) in water (7 mL), and the mixture was stirredat ambient temperature for 1 hour. Acetic acid (0.45 mL, 7.9 mmol) wasslowly added and the mixture was evaporated to dryness and purified byflash chromatography (100 g silica cartridge) using a gradient elutionfrom 0% to 15% methanol in dichloromethane (+1% triethylamine) to affordthe title compound (1.37 g, 70% yield). LCMS RT=0.99 min, ES+ve m/z 399[M+H]⁺.

tert-butyl((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)carbamate

A stirred mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (125 mg, 0.35 mmol) and(S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid (commerciallyavailable from for example Aldrich) (77 mg, 0.35 mmol) in DMF (0.9 mL)was treated with DIPEA (0.22 mL, 1.3 mmol) and then with HATU (134 mg,0.35 mmol) and the mixture was stirred at ambient temperature for 1hour. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (120 mg, 72% yield). LCMS RT=0.87 min, ES+ve m/z 517 [M+H]⁺.

Using a method analogous to that fortert-butyl-((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)carbamate, the following compounds were prepared:

Name Structure Yield RT [M + H]+ (S)-tert-butyl 2-((2S,4R)-4-hydroxy-2-((4-(4- methylthiazol-5- yl)benzyl)carbamoyl) pyrrolidine-1-carbonyl)piperidine-1- carboxylate

67% 0.88 min 529 (S)-tert-butyl 3-((2S,4R)- 4-hydroxy-2-((4-(4-methylthiazol-5- yl)benzyl)carbamoyl) pyrrolidine-1-carbonyl)morpholine-4- carboxylate

67% 0.78 min 531 tert-butyl ((S)-1-((2S,4R)- 4-hydroxy-2-((4-(4-methylthiazol-5- yl)benzyl)carbamoyl) pyrrolidin-1-yl)-1-oxobutan-2-yl)carbamate

85% 0.81 min 503 tert-butyl ((S)-1-((2S,4R)- 4-hydroxy-2-((4-(4-methylthiazol-5- yl)benzyl)carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2- yl)carbamate

85% 0.94 min 531 tert-butyl ((S)-1- cyclopropyl-2-((2S,4R)-4-hydroxy-2-((4-(4- methylthiazol-5- yl)benzyl)carbamoyl)pyrrolidin-1-yl)-2- oxoethyl)carbamate

82% 0.83 min 515

(2S,4R)-1-(2-aminoacetyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (100 mg, 0.28 mmol) and2-((tert-butoxycarbonyl)amino)acetic acid (commercially available fromfor example Aldrich) (49 mg, 0.28 mmol) in DMF (3 mL) was treated withDIPEA (0.20 mL, 1.1 mmol) and then with HATU (118 mg, 0.31 mmol) and themixture was stirred at ambient temperature for 30 minutes. Water (20 ml)was added and the product was extracted with ethyl acetate (3×20 mL).The combined organic phase was washed with saturated aqueous sodiumbicarbonate (20 mL), water (20 mL), brine (20 mL) filtered through ahydrophobic frit and evaporated to dryness. The residue was thendissolved in dichloromethane (3 mL) and treated with TFA (1 mL, 13mmol). After stirring at ambient temperature for 10 minutes, thereaction mixture was evaporated to dryness. The residue was dissolved inthe minimum amount of methanol and then loaded onto a pre-conditioned(methanol) aminopropyl solid-phase extraction cartridge (5 g). Thecolumn was eluted with methanol (3 volumes) and the product-containingfractions were evaporated to dryness to afford the title compound (104mg, 99% yield). LCMS RT=0.44 min, ES+ve m/z 375 [M+H]⁺.

(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1-((S)-morpholine-3-carbonyl)pyrrolidine-2-carboxamide

A solution of (S)-tert-butyl3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)morpholine-4-carboxylate(115 mg, 0.22 mmol) in dichloromethane (0.5 mL) was treated with TFA(0.5 mL) and the reaction mixture was stirred at ambient temperature for1 hour. The mixture was evaporated to dryness and the residue was thendissolved in the minimum amount of a mixture of methanol:dichloromethane(1:1), and loaded onto a pre-conditioned (methanol) aminopropylsolid-phase extraction cartridge (2 g). The column was eluted withmethanol (3 volumes) and the product-containing fractions wereevaporated under reduced pressure to afford the title compound (89 mg,94% yield). LCMS RT=0.47 min, ES+ve m/z 431 [M+H]⁺.

(2S,4R)-4-hydroxy-1-((S)-3-methyl-2-(methylamino)butanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride

A mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (100 mg, 0.28 mmol),(S)-2-((tert-butoxycarbonyl)(methyl)amino)-3-methylbutanoic acid(commercially available from for example Aldrich) (65 mg, 0.28 mmol) andDIPEA (0.247 mL, 1.41 mmol) in DMF (2 mL) was treated with HATU (118 mg,0.31 mmol) and stirred at ambient temperature for 30 minutes. The Bocprotected intermediate was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier). The purifiedintermediate was dissolved in methanol:dichloromethane (1:1, 3 mL),treated with hydrochloric acid in 1,4-dioxane (4M, 3 mL, 12 mmol) andallowed to stand for 1 hour. The mixture was then evaporated to drynessto afford the title compound (107 mg, 0.23 mmol, 81% yield). LCMSRT=0.55 min, ES+ve m/z 431 [M+H]⁺.

(2S,4R)-1-((S)-2-amino-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride

A solution of tert-butyl((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)carbamate(287 mg, 0.56 mmol) in THF (5 mL) and treated with 4M hydrochloric acidin 1,4-dioxan (10 mL) and stirred at ambient temperature for 2 hours.The mixture was evaporated to dryness to afford the title compound (224mg, 0.49 mmol, quantitative). LCMS RT=0.55 min, ES+ve m/z 417 [M+H]⁺.

(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride

A stirred mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (70 mg, 0.20 mmol) and(S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoic acid(commercially available from for example Fluka) (50 mg, 0.22 mmol) inDMF (1 mL) was treated with DIPEA (0.14 mL, 0.79 mmol) and then withHATU (90 mg, 0.24 mmol), and stirred at ambient temperature for 30minutes. The product was subjected to purification by mass-directedautomated preparative HPLC (formic acid modifier) to give theintermediate boc-protected product. The intermediate was then dissolvedin a mixture of dichloromethane (0.5 mL) and methanol (0.1 mL) andtreated with 4M hydrochloric acid in 1,4-dioxane (0.25 mL, 1.0 mmol),After stirring at ambient temperature for 1 hour, the reaction mixturewas evaporated to dryness and the residue triturated to a solid withdichloromethane and dried under vacuum to afford the title compound (76mg, 82% yield). LCMS RT=0.58 min, ES+ve m/z 431 [M+H]⁺.

(2S,4R)-1-((S)-2-((S)-2-amino-4-methylpentanamido)propanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride

A solution of a mixture of(2S,4R)-1-((S)-2-aminopropanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (507 mg, 1.2 mmol), DIPEA (0.868 mL, 4.97 mmol) and(S)-2-((tert-butoxycarbonyl)amino)-4-methylpentanoic acid (commerciallyavailable from for example Aldrich) (230 mg, 0.99 mmol) in DMF (5 mL)was treated with HATU (416 mg, 1.1 mmol) and stirred at ambienttemperature for 2 hours. Water (50 mL) was added and the product wasextracted with ethyl acetate (50 mL). The organic phase was washed withbrine (2×50 mL), dried using a hydrophobic frit and evaporated todryness. The residue was dissolved in methanol:dichloromethane (1:1, 15mL), treated with hydrochloric acid in 1,4-dioxane (4M, 5 mL, 20 mmol)and stirred at ambient temperature for 3 hours. The mixture wasevaporated to dryness and the residue was suspended in dichloromethane(10 mL), sonicated, filtered and dried under vacuum to afford the titlecompound (280 mg, 0.56 mmol, 56% yield). LCMS RT=0.55 min, ES+ve m/z 502[M+H]⁺.

(2S,4R)-tert-butyl2-((4-(2,4-dimethylthiazol-5-yl)benzyl)carbamoyl)-4-hydroxypyrrolidine-1-carboxylate

Under an atmosphere of nitrogen, a mixture of (2S,4R)-tert-butyl2-((4-bromobenzyl)carbamoyl)-4-hydroxypyrrolidine-1-carboxylate (200 mg,0.50 mmol), 2,4-dimethylthiazole (commercially available from forexample Avocado) (113 mg, 1.0 mmol), palladium(II) acetate (commerciallyavailable from for example Aldrich) (2 mg, 10 μmol) and potassiumacetate (98 mg, 1.0 mmol) in N-methyl-2-pyrrolidone (2 mL) was stirredat 120° C. for 18 hours. The cooled mixture was treated with water (25ml) and the product was extracted with ethyl acetate (4×30 mL). Thecombined organic phase was washed with brine (5×20 mL), filtered througha hydrophobic frit and evaporated to dryness. The product was subjectedto purification by mass-directed automated preparative HPLC (formic acidmodifier) to afford the title compound (142 mg, 66% yield). LCMS RT=0.77min, ES+ve m/z 432 [M+H]⁺.

(2S,4R)—N-(4-(2,4-dimethylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide,hydrochloride

A solution of (2S,4R)-tert-butyl2-((4-(2,4-dimethylthiazol-5-yl)benzyl)carbamoyl)-4-hydroxypyrrolidine-1-carboxylate(142 mg, 0.33 mmol) in a mixture of methanol (0.5 mL) anddichloromethane (1.5 mL) was treated with 4M hydrochloric acid in1,4-dioxane (0.63 mL, 2.5 mmol) and stirred at ambient temperature for 2hours. The solvent was evaporated to dryness and the residue wastriturated to a solid with diethyl ether and dried under vacuum toafford the title compound (120 mg, 99% yield). LCMS RT=0.49 min, ES+vem/z 332 [M+H]⁺.

(S)-3-methyl-2-(3-methyl-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)butanoicacid

A mixture of 3-methylfuran-2,5-dione (commercially available from forexample Aldrich) (0.12 mL, 1.3 mmol) and (S)-2-amino-3-methylbutanoicacid (commercially available from for example Apollo Scientific) (150mg, 1.3 mmol). in acetic acid (1 mL) was sealed and heated in a Biotage“Initiator” microwave at 120° C. for 1 hour. The mixture was evaporatedto dryness to afford the title compound (253 mg, 94% yield). LCMSRT=0.75 min, ES+ve m/z 212 [M+H]⁺

benzyl 2-(3-oxomorpholino)propanoate

Under an atmosphere of nitrogen, an ice-cooled solution ofmorpholin-3-one (commercially available from for example Aldrich) (100mg, 1.0 mmol) in DMF (2 mL) was treated with sodium hydride (60% w/w inmineral oil) (40 mg, 1.0 mmol). After 5 minutes, benzyl2-bromopropanoate (commercially available from for example Aldrich) (240mg, 1.0 mmol) was added and the mixture was stirred with cooling for 30minutes and then at ambient temperature for a further 18 hours. Thereaction mixture was cautiously treated with saturated aqueous sodiumbicarbonate (10 mL) and then extracted with ethyl acetate (2×40 mL). Thecombined organic phase was washed with brine (25 mL), filtered through ahydrophobic frit, and evaporated to dryness. The product was subjectedto purification by mass-directed automated preparative HPLC (formic acidmodifier) to give the title compound (105 mg, 40% yield). LCMS RT=0.80min, ES+ve m/z 264 [M+H]⁺.

2-(3-oxomorpholino)propanoic acid

Under an atmosphere of nitrogen, a solution of benzyl2-(3-oxomorpholino)propanoate (90 mg, 0.34 mmol) in ethanol (3 mL) wasadded to a flask containing palladium on carbon (36 mg, 0.034 mmol)(10%, Degussa type). The flask was then filled with hydrogen and themixture was stirred at ambient temperature for 1 hour. The catalyst wasremoved by filtration through celite and the filtrate was evaporatedunder reduced pressure to afford the title compound (55 mg, 93% yield).LCMS RT=0.33 min, ES+ve m/z 174 [M+H]⁺. tert-butyl4-(1-(benzyloxy)-1-oxopropan-2-yl)-3-oxopiperazine-1-carboxylate

Under an atmosphere of nitrogen, an ice-cooled solution of tert-butyl3-oxopiperazine-1-carboxylate (commercially available from for exampleAldrich) (200 mg, 1.0 mmol) in DMF (4 mL) was treated with sodiumhydride (60% w/w in mineral oil) (44 mg, 1.1 mmol). After 5 minutes,benzyl 2-bromopropanoate (commercially available from for exampleAldrich) (255 mg, 1.05 mmol) was added. The mixture was stirred at 0° C.for 30 minutes and then at ambient temperature for a further 18 hours.The mixture was cautiously treated with saturated aqueous sodiumbicarbonate (20 mL) and then extracted with ethyl acetate (2×40 mL). Thecombined organic phase was washed with brine (25 mL), filtered through ahydrophobic frit, and evaporated to dryness. The product was purified byflash column chromatography (20 g silica cartridge) using a gradientelution from 0% to 100% ethyl acetate in cyclohexane to afford the titlecompound (230 mg, 64% yield). LCMS RT=1.05 min, ES+ve m/z 363 [M+H]⁺.

2-(4-(tert-butoxycarbonyl)-2-oxopiperazin-1-yl)propanoic acid

Under an atmosphere of nitrogen, a solution of tert-butyl4-(1-(benzyloxy)-1-oxopropan-2-yl)-3-oxopiperazine-1-carboxylate (230mg, 0.64 mmol) in ethanol (3 mL) was added to a flask containingpalladium on carbon (68 mg, 0.063 mmol) (10%, Degussa type). The flaskwas filled with hydrogen and the mixture was stirred at ambienttemperature for 1 hour. The catalyst was removed by filtration throughcelite and the filtrate was evaporated under reduced pressure to affordthe title compound (171 mg, 99% yield). LCMS RT=0.62 min, ES+ve m/z 290[M+H]⁺.

tert-butyl 4-(2-oxo-2,3-dihydrobenzo [d]oxazol-5-yl)benzylcarbamate

Under an atmosphere of nitrogen, a mixture of(4-(((tert-butoxycarbonyl)amino)methyl)phenyl)boronic acid (commerciallyavailable from for example Aldrich) (387 mg, 1.54 mmol),5-bromobenzo[d]oxazol-2(3H)-one (commercially available from for exampleAldrich) (300 mg, 1.40 mmol) and sodium carbonate (446 mg, 4.21 mmol) inDMF (4 mL) was treated withdicholoro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)(commercially available from for example Aldrich) (72 mg, 0.098 mmol)then sealed and heated in a Biotage “Initiator” microwave at 110° C. for1 hr. The cooled product mixture was treated with dichloromethane (50mL) and water (10 mL). The mixture was separated and the organicfraction was evaporated to dryness. The product was subjected topurification by mass-directed automated preparative HPLC (formic acidmodifier) to afford the title compound (178 mg, 0.52 mmol, 37% yield).LCMS RT=1.03 min, ES+ve m/z 341 [M+H]⁺.

5-(4-(aminomethyl)phenyl)benzo[d]oxazol-2(3H)-one, hydrochloride

A solution of tert-butyl4-(2-oxo-2,3-dihydrobenzo[d]oxazol-5-yl)benzylcarbamate (130 mg, 0.38mmol) in THF (10 mL) was treated with hydrochloric acid (4M in1,4-dioxan) (1 mL, 4 mmol) and the mixture was stirred at ambienttemperature overnight. The mixture was treated with diethyl ether (40mL); the resulting precipitate was filtered off and dried under vacuumto afford the title compound (87 mg, 0.31 mmol, 82% yield). LCMS RT=0.49min, ES+ve m/z 241 [M+H]⁺.

(2S,4R)-4-hydroxy-1-(3-hydroxybenzoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

An ice-cooled solution of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (217 mg, 0.61 mmol), 3-hydroxybenzoic acid (85 mg, 0.61mmol) and DIPEA (0.321 mL, 1.84 mmol) in DMF (4 mL) was treatedportion-wise with HATU (240 mg, 0.63 mmol) over 1 minute and thenstirred at ambient temperature for 1 hour. The mixture was treated withsaturated aqueous sodium bicarbonate (20 mL) and extracted with ethylacetate (3×20 mL). The combined organic phase was washed with brine(3×30 mL), filtered through a hydrophobic frit and then evaporated todryness. The product was purified by flash chromatography (50 g silicacartridge) using a gradient elution from 0 to 25% methanol indichloromethane to afford the title compound (234 mg, 0.53 mmol, 87%yield). LCMS RT=0.49 min, ES+ve m/z 241 [M+H]⁺.

(S)-2-(1-oxoisoindolin-2-yl)propanoic acid

A mixture of phthalaldehyde (commercially available from for exampleAldrich) (4 g, 30 mmol) and (S)-2-aminopropanoic acid (commerciallyavailable from for example Aldrich) (2.39 g, 27 mmol) in acetonitrile(150 mL) was heated at reflux for 5 hr then allowed to cool to ambienttemperature and stood overnight. The resulting crystalline precipitatewas filtered off, washed with acetonitrile and dried under vacuum toafford the title compound (4.46 g, 22 mmol, 73% yield). LCMS RT=0.59min, ES+ve m/z 206 [M+H]⁺.

Using a method analogous to that for(S)-2-(1-oxoisoindolin-2-yl)propanoic acid, the following compounds wereprepared:

Name Structure Yield RT [M + H]+ (S)-4-methoxy-2-(1- oxoisoindolin-2-yl)butanoic acid

46% 0.62 min 250 (S)-2-(1-oxoisoindolin-2- yl)buanoic acid

60% 0.67 min 220 (S)-2-(1-oxoisoindolin-2- yl)pentanoic acid

60% 0.77 min 234 (S)-2-cyclopropyl-2-(1- oxoisoindolin-2-yl)acetic acid

54% 0.69 min 232 (S)-3-methyl-2-(1- oxoisoindolin-2- yl)butanoic acid

59% 0.74 min 234 (S)-3,3-dimethyl-2-(1- oxoisoindolin-2- yl)butanoicacid

69% 0.82 min 248

Ethyl 2-(5-methoxy-1-oxoisoindolin-2-yl)-3-methylbutanoate

Under an atmosphere of nitrogen, an ice-cooled solution of5-methoxyisoindolin-1-one (commercially available from for example ChemImpex) (105 mg, 0.64 mmol) in DMF (2.5 mL) was treated with sodiumhydride (60% w/w in mineral oil) (31 mg, 0.77 mmol). The mixture waswarmed to ambient temperature, treated with ethyl2-bromo-3-methylbutanoate (commercially available from for example AlfaAesar) (135 mg, 0.64 mmol), stirred at ambient temperature for 2 hoursand then heated to 70° C. for a further 18 hours. The mixture was thenice-cooled and treated with additional sodium hydride (60% w/w inmineral oil) (31 mg, 0.77 mmol), followed by ethyl2-bromo-3-methylbutanoate (135 mg, 0.64 mmol) and stirred at 70° C. fora further 24 hours. The cooled mixture was then cautiously treated withsaturated aqueous ammonium chloride (20 mL) and the product wasextracted with ethyl acetate (2×25 mL). The combined organic phase waswashed with water (20 mL), brine (20 mL), filtered through a hydrophobicfrit and evaporated to dryness. The product was purified by flashchromatography (20 g silica cartridge) using a gradient elution from 0%to 50% ethyl acetate in cyclohexane to afford the title compound (44 mg,23% yield). LCMS RT=1.01 min, ES+ve m/z 292 [M+H]⁺

Ethyl 2-(6-methoxy-1-oxoisoindolin-2-yl)-3-methylbutanoate

Under an atmosphere of nitrogen, an ice-cooled solution of6-methoxyisoindolin-1-one (commercially available from for exampleAstatech) (105 mg, 0.64 mmol) in DMF (2.5 mL) was treated with sodiumhydride (60% w/w in mineral oil) (31 mg, 0.77 mmol) and the mixture wasallowed to warm to ambient temperature. The mixture was then treatedwith ethyl 2-bromo-3-methylbutanoate (commercially available from forexample Alfa Aesar) (135 mg, 0.64 mmol) and the mixture was stirred for18 hours then cautiously treated with saturated aqueous ammoniumchloride (20 mL). The product was extracted with ethyl acetate (2×25 mL)and the combined organic phase was washed with water (20 mL), brine (20mL), filtered through a hydrophobic frit and evaporated to dryness. Theproduct was purified by flash chromatography (20 g silica cartridge)using a gradient elution from 0 to 50% ethyl acetate in cyclohexane toafford the title compound (40 mg, 21% yield). LCMS RT=1.03 min, ES+vem/z 292 [M+H]⁺

2-(6-methoxy-1-oxoisoindolin-2-yl)-3-methylbutanoic acid

A solution of ethyl 2-(6-methoxy-1-oxoisoindolin-2-yl)-3-methylbutanoate(40 mg, 0.14 mmol) in ethanol (0.4 mL) was treated with 2M aqueoussodium hydroxide (0.20 mL, 0.41 mmol) and the mixture was stirred atambient temperature for 2 hours. The reaction mixture was evaporated todryness, treated with water (10 mL) and acidified to pH 3 using 2Maqueous hydrochloric acid. The product was extracted with ethyl acetate(2×10 mL), and the combined organic phase was filtered through ahydrophobic frit and evaporated to dryness to afford the title compound(34 mg, 94% yield). LCMS RT=0.80 min, ES+ve m/z 264 [M+H]⁺.

2-(5-methoxy-1-oxoisoindolin-2-yl)-3-methylbutanoic acid

A solution of ethyl2-(5-methoxy-1l-oxoisoindolin-2-yl)-3-methylbutanoate (40 mg, 0.14 mmol)in ethanol (0.4 mL) was treated with 2M aqueous sodium hydroxide (0.20mL, 0.41 mmol) and the mixture was stirred at ambient temperature for 2hours. The mixture was then evaporated to dryness; the residue wastreated with water (10 mL) and acidified to pH 3 using 2M aqueoushydrochloric acid. The product was extracted with ethyl acetate (2×10mL), and the combined organic phase was filtered through a hydrophobicfrit and evaporated to dryness to afford the title compound (33 mg, 93%yield). LCMS RT=0.76 min, ES+ve m/z 264 [M+H]⁺.

Ethyl 2-(7-chloro-1-oxoisoindolin-2-yl)-3-methylbutanoate

Under a nitrogen atmosphere, an ice-cooled solution of7-chloroisoindolin-1-one (commercially available from for example JWPharm) (150 mg, 0.90 mmol) in DMF (2.5 mL) was treated with sodiumhydride (60% w/w in mineral oil) (50 mg, 1.25 mmol). The mixture wasallowed to warm to ambient temperature, then treated with ethyl2-bromo-3-methylbutanoate (commercially available from for example AlfaAesar) (187 mg, 0.90 mmol) and stirred for 5 hours. The mixture was thenice-cooled, treated with additional sodium hydride (60% w/w in mineraloil) (50 mg, 1.25 mmol) and ethyl 2-bromo-3-methylbutanoate (187 mg,0.90 mmol) and the mixture was stirred at ambient temperature for afurther 24 hours. The mixture was then cautiously treated with saturatedaqueous ammonium chloride (20 mL), and the product was extracted withethyl acetate (2×25 mL). The combined organic phase was washed withwater (20 mL), brine (20 mL), filtered through a hydrophobic frit andevaporated to dryness. The product was purified by flash chromatography(20 g silica cartridge) using a gradient elution from 0 to 50% ethylacetate in cyclohexane to afford the title compound (40 mg, 15% yield).LCMS RT=1.07 min, ES+ve m/z 296 [M+H]⁺.

2-(7-chloro-1-oxoisoindolin-2-yl)-3-methylbutanoic acid

A solution of ethyl 2-(7-chloro-1-oxoisoindolin-2-yl)-3-methylbutanoate(40 mg, 0.14 mmol) in ethanol (0.4 mL) was treated with 2M aqueoussodium hydroxide (0.22 mL, 0.45 mmol) and the mixture was stirred atambient temperature for 2 hours. The mixture was then evaporated todryness, treated with water (10 mL) and then acidified to pH 3 using 2Maqueous hydrochloric acid. The product was extracted with ethyl acetate(2 xl0 mL) and the combined organic phase was filtered through ahydrophobic frit and evaporated to dryness to afford the title compound(34 mg, 94% yield). LCMS RT=0.82 min, ES+ve m/z 268 [M+H]⁺.

2-Hydroxy-4-(4-methylthiazol-5-yl)benzonitrile

Under an atmosphere of nitrogen, a mixture of4-bromo-2-hydroxybenzonitrile (commercially available from for exampleFluorochem) (15 g, 76 mmol), 4-methylthiazole (commercially availablefrom for example Aldrich) (14 mL, 152 mmol), potassium acetate (14.9 g,152 mmol) and palladium(II) acetate (0.34 g, 1.52 mmol) in1-methyl-2-pyrrolidone (125 mL) was heated at 110° C. for 3 hours. Themixture was then cooled to 50° C., poured into water (300 mL) andextracted with ethyl acetate (3×350 mL). The combined organic fractionwas filtered and the filtrate was then washed with brine (3×400 mL),filtered through a hydrophobic frit and evaporated to dryness. Theresidue was re-evaporated from toluene then from diethyl ether and thenslurried in methanol to precipitate a yellow solid which was filteredoff. The filtrate was evaporated to dryness and slurried in ice-cooledmethanol to afford a second batch of yellow solid. The combined solidwas dried under vacuum to afford the title compound (12 g, 56 mmol, 73%yield). LCMS RT=0.75 min, ES+ve m/z 217 [M+H]⁺.

2-(Aminomethyl)-5-(4-methylthiazol-5-yl)phenol

An ice-cooled solution of 2-hydroxy-4-(4-methylthiazol-5-yl)benzonitrile(12 g, 56 mmol) in THF (550 mL) was treated dropwise with lithiumaluminium hydride (1M in THF, 140 mL, 140 mmol) over 5 minutes. Theresulting mixture was then heated at 50° C. for 30 minutes andadditional lithium aluminium hydride (1M in THF, 20 mL, 20 mmol) wasadded. After a further 30 minutes the mixture was cooled in an ice bathand treated cautiously with water (14 mL), followed by aqueous sodiumhydroxide (4M, 42 mL, 168 mmol) and finally water (14 mL). Afterstanding for 3 days, the mixture was filtered and the filtered solid waswashed with THF. The combined filtrate was evaporated to dryness and theresidue was slurried in dichloromethane:methanol (4:1) with Celite(about 20 g) and filtered. The filtered solid was washed three timeswith dichloromethane/methanol (4:1) and the combined filtrate wasevaporated to dryness. The product was purified by flash chromatography(330 g silica cartridge) using a gradient elution from 0 to 15% methanolin dichloromethane (+1% triethylamine) to afford the title compound (6.2g, 28 mmol, 51% yield). LCMS RT=0.41 min, ES+ve m/z 221 [M+H]⁺.

(2S,4R)-tert-Butyl4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate

An ice-cooled solution of 2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol(3.05 g, 13.8 mmol) and(2S,4R)-1-(tert-butoxycarbonyl)-4-hydroxypyrrolidine-2-carboxylic acid(2.94 mL, 13.8 mmol) in DMF (35 mL) was treated with DIPEA (7.25 mL, 42mmol) followed by HATU (5.79 g, 15.2 mmol) and the mixture was stirredat ambient temperature for 1 hour. The mixture was treated withsaturated aqueous sodium bicarbonate (50 mL) and extracted with ethylacetate (3×80 mL). The combined organic phase was washed with brine (60mL), filtered through a hydrophobic frit and evaporated to dryness. Theproduct was purified by flash chromatography (330 g silica cartridge)using a gradient from 0 to 15% methanol in dichloromethane to afford thetitle product (4.8 g, 11 mmol, 80% yield). LCMS RT=0.76 min, ES+ve m/z434 [M+H]⁺.

(2S,4R)-4-Hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride

A solution of (2S,4R)-tert-butyl4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate(4.8 g, 11 mmol) in dichloromethane:methanol 20:1 (50 mL) was treatedwith hydrochloric acid (4M in 1,4-dioxane) (35 mL, 140 mmol) and themixture was stirred overnight at ambient temperature. The mixture wasthen evaporated to dryness and the residual solid was suspended indichloromethane and filtered. The filtered solid was washed with furtherdichloromethane and dried under vacuum to afford the title compound (4g, 10.8 mmol, 98% yield). LCMS RT=0.46 min, ES+ve m/z 334 [M+H]⁺.

(2S,4R)-1-((S)-2-Aminopropanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride

A stirred mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (100 mg, 0.28 mmol),(S)-2-((tert-butoxycarbonyl)amino)propanoic acid (commercially availablefrom for example Aldrich) (64 mg, 0.34 mmol) and DIPEA (0.197 mL, 1.13mmol) in dry DMF (3 mL) was treated with HATU (129 mg, 0.34 mmol andstirred at ambient temperature for 30 minutes. The mixture was thenpartitioned between ethyl acetate (30 mL) and water (30 mL) and theorganic phase was washed with brine (30 mL), dried (hydrophobic frit)and evaporated to dryness. The residue was dissolved in methanol andadded to a methanol-preconditioned aminopropyl solid-phase extractioncartridge (2 g) eluting with methanol (3 column volumes). The resultingeluant was evaporated to dryness and the residue was dissolved indichloromethane:methanol (1:1, 8 mL) and treated with hydrochloric acid,(4M in 1,4-dioxane) (1 mL, 4 mmol). The mixture was stirred at ambienttemperature for 16 hours and then evaporated to dryness to afford thetitle compound (107 mg, 0.25 mmol, 89% yield). LCMS RT=0.51 min, ES+vem/z 389 [M+H]⁺.

(2S,4R)-1-(2-Amino-2-methylpropanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A solution of a mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (100 mg, 0.28 mmol),2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (commerciallyavailable from for example Aldrich) (69 mg, 0.34 mmol) and DIPEA (0.197mL, 1.13 mmol) in dry DMF (3 mL) was treated with HATU (129 mg, 0.34mmol, stirred at ambient temperature for 30 minutes and then partitionedbetween ethyl acetate (30 mL) and water (30 mL). The organic phase waswashed with brine (30 mL), dried (hydrophobic frit) and evaporated todryness. The residue was dissolved in methanol and added to amethanol-preconditioned aminopropyl solid-phase extraction cartridge (2g) eluting with methanol. The resulting eluant was evaporated to drynessand the residue was dissolved in dichloromethane:methanol (1:1, 8 mL)and treated with hydrochloric acid, 4M in 1,4-dioxane (1 mL, 4 mmol).The reaction mixture was stirred at ambient temperature for 16 hours andthen evaporated to dryness. The residue was suspended in dichloromethane(4 mL) and treated with TFA (1 mL) and the mixture was stirred atambient temperature for 4 hours.

The mixture was evaporated to dryness and the residue was dissolved inmethanol and added to a methanol-preconditioned sulfonic acidsolid-phase extraction cartridge (2 g) and eluted with methanol (3column volumes) and then with ammonia in methanol (2M, 3 columnvolumes). The product-containing fractions were evaporated to dryness toafford the title compound (95 mg, 0.24 mmol, 84% yield). LCMS RT=0.53min, ES+ve m/z 403 [M+H]⁺.

(2S,4R)-4-Hydroxy-1-((S)-2-(methylamino)propanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

A stirred mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (120 mg, 0.34 mmol) and(S)-2-((tert-butoxycarbonyl)(methyl)amino)propanoic acid (commerciallyavailable from for example Aldrich) (76 mg, 0.37 mmol) in DMF (2 mL) wastreated with DIPEA (0.24 mL, 1.36 mmol) and then with HATU (155 mg, 0.41mmol), and the mixture was stirred at ambient temperature for 30minutes. The crude product was subjected to purification bymass-directed automated preparative HPLC (formic acid modifier) to givethe intermediate Boc-protected product. The intermediate was suspendedin dichloromethane (0.5 mL) and treated with TFA (0.5 mL). The mixturewas stirred at ambient temperature for 1 hour and was then evaporated todryness. The residue was dissolved in the minimum amount of a mixture ofmethanol:dichloromethane (1:1), and then loaded onto a pre-conditioned(methanol) aminopropyl solid-phase extraction cartridge (5 g). Thecolumn was eluted with methanol (3 volumes) and the product-containingfractions were combined and evaporated to dryness to afford the titlecompound (103 mg, 75% yield). LCMS RT=0.47 min, ES+ve m/z 403 [M+H]⁺.

(2S,4R)-4-Hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)-1-((S)-morpholine-3-carbonyl)pyrrolidine-2-carboxamide,hydrochloride

A mixture of(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (100 mg, 0.28 mmol),(S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (commerciallyavailable from for example Astatech) (65 mg, 0.28 mmol) and DIPEA (0.247mL, 1.41 mmol) in DMF (2 mL) was treated with HATU (118 mg, 0.311 mmol)and stirred at ambient temperature for 30 minutes. The Boc protectedintermediate was subjected to purification by mass-directed automatedpreparative HPLC (formic acid modifier). The intermediate was dissolvedin methanol:dichloromethane (1:1, 3 mL), treated with hydrochloric acidin 1,4-dioxane (4M, 3 mL, 12 mmol) and allowed to stand for 1 hour. Themixture was evaporated to dryness to afford the title compound (110 mg,0.24 mmol, 83% yield). LCMS RT=0.50 min, ES+ve m/z 431/432 [M+H]⁺.

Tert-butyl4-(3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)phenoxy)butanoate

A solution of a mixture of 3-(4-(tert-butoxy)-4-oxobutoxy)benzoic acid(95 mg, 0.34 mmol),(2S,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (100 mg, 0.28 mmol) and DIPEA (0.2 mL, 1.15 mmol) in dryDMF (3 mL) was treated with HATU (129 mg, 0.34 mmol) and the mixture wasstirred at ambient temperature for 30 minutes. The mixture was added toan aminopropyl solid-phase extraction cartridge and eluted with methanol(3 column volumes). The resulting eluant was evaporated to dryness andthe product was subjected to purification by mass-directed automatedpreparative HPLC (formic acid modifier) to afford the title compound(130 mg, 0.22 mmol, 79% yield). LCMS RT=0.98 min, ES+ve m/z 580 [M+H]⁺.

Example—VHL Protac Which Binds Estrogen Receptor (Estrogen Protacts)Abbreviations

-   DCM: dichloromethane.-   DIPEA: N,N-diisopropylethylamine.-   DMF: N,N-dimethylformamide.-   HATU: 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium    hexafluorophosphate.-   HPLC: high-performance liquid chromatography.-   LCMS: liquid chromatography-mass spectrometry-   Min: minutes.-   RT: retention time.-   tBu: tert-butoxide.-   TFA: trifluoroacetic acid.-   THF: tetrahydrofuran.

LCMS Method:

The analysis was conducted on an Acquity UPLC BEH C18 column (50 mm×2.1mm internal diameter 1.7 μm packing diameter) at 40° C.

The solvents employed were:

A=0.1% v/v solution of formic acid in water.B=0.1% v/v solution of formic acid in acetonitrile.

The gradient employed was as follows:

Time Flow Rate (minutes) (mL/min) % A % B 0 1 97 3 1.5 1 0 100 1.9 1 0100 2.0 1 97 3

The UV detection was an averaged signal from wavelength of 210 nm to 350nm and mass spectra were recorded on a mass spectrometer usingalternate-scan positive and negative mode electrospray ionization.

The following illustrates the mobile phases and gradients used whencompounds underwent purification by mass-directed autopreparative HPLC.

Mass-Directed Autopreparative HPLC (Formic Acid Modifier)

The HPLC analysis was conducted on a Sunfire C18 column (150 mm×30 mminternal diameter, 5 m packing diameter) at ambient temperature.

The solvents employed were:

A=0.1% v/v solution of formic acid in water.B=0.1% v/v solution of formic acid in acetonitrile.

Mass-Directed Autopreparative HPLC (Trifluoroacetic Acid Modifier)

The HPLC analysis was conducted on a Sunfire C18 column (150 mm×30 mminternal diameter, 5 m packing diameter) at ambient temperature.

The solvents employed were:

A=0.1% v/v solution of trifluoroacetic acid in water.B=0.1% v/v solution of trifluoroacetic acid in acetonitrile.

Mass-Directed Autopreparative HPLC (Ammonium Bicarbonate Modifier)

The HPLC analysis was conducted on an XBridge C18 column (150 mm×30 mminternal diameter, 5 m packing diameter) at ambient temperature.

The solvents employed were:

A=10 mM ammonium bicarbonate in water adjusted to pH 10 with ammoniasolution.B=acetonitrile.

For each of the mass-directed autopreparative purifications,irrespective of the modifier used, the gradient employed was dependentupon the retention time of the particular compound undergoingpurification as recorded in the analytical LCMS, and was as follows:

For compounds with an analytical LCMS retention time below 0.6 minutesthe following gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B 0 40 99 1 1 40 99 1 10 40 7030 11 40 1 99 15 40 1 99

For compounds with an analytical LCMS retention time between 0.6 and 0.9minutes the following gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B 0 40 85 15 1 40 85 15 10 40 4555 11 40 1 99 15 40 1 99

For compounds with an analytical LCMS retention time between 0.9 and 1.2minutes the following gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B  0 40 70 30  1 40 70 30 10 4015 85 11 40 1 99 15 40 1 99

For compounds with an analytical LCMS retention time between 1.2 and 1.4minutes the following gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B 0 40 50 50 1 40 50 50 10 40 199 11 40 1 99 15 40 1 99

For compounds with an analytical LCMS retention time greater than 1.4minutes (LCMS method A) or greater than 3.6 minutes (LCMS method B) thefollowing gradient was used:

Time Flow Rate (minutes) (mL/min) % A % B 0 40 20 80 1 40 20 80 10 40 199 11 40 1 99 15 40 1 99

The UV detection was an averaged signal from wavelength of 210 nm to 350nm and mass spectra were recorded on a mass spectrometer usingalternate-scan positive and negative mode electrospray ionization.

The chemical names were generated using ACD Name Pro version 6.02 fromAdvanced Chemistry Development, Inc.

8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one

(8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-onecan be prepared according to the process described by Xiang-Rong Jiang,J. Walter Sowell, Bao Ting Zhu, Steroids, 2006, 71, 334-342.(doi:10.1016/j.steroids. 2005.11.008).

15-Bromo-1-phenyl-2,5,8,11-tetraoxapentadecane

To a suspension of sodium hydride, 60% w/w in mineral oil (0.250 g, 6.24mmol) in DMF (2 mL) was added a solution of2-(2-(2-(benzyloxy)ethoxy)ethoxy)ethanol (1 g, 4.16 mmol) (commerciallyavailable from for example Fluorochem) in DMF (2 mL) at 0° C. Afterstirring for 25 minutes, 1,4-dibromobutane (commercially available fromfor example Aldrich) (4.04 g, 18.73 mmol) dissolved in DMF (2 mL) wasadded dropwise to the mixture. The reaction was stirred under anatmosphere of nitrogen for 2.5 hours. A further aliquot of sodiumhydride, 60% w/w in mineral oil (0.250 g, 6.24 mmol) was added and thereaction was stirred at 0° C. for 30 minutes. The reaction was warmed toroom temperature and stirred for 30 minutes. A final aliquot of sodiumhydride, 60% w/w in mineral oil (0.250 g, 6.24 mmol) was added and thereaction stirred at room temperature for 2 hours then left standing overthe weekend. The reaction mixture was filtered through celite and thesolid washed with DCM. The filtrate was partitioned between DCM (30 mL)and water (30 mL). The organic extract was washed with brine (2×30 mL),dried using a hydrophobic frit and concentrated under reduced pressure.The product was purified by chromatography on silica using a gradientelution from 0% to 100% methyl tert-butyl ether in cyclohexane to affordthe title compound (711 mg, 1.89 mmol, 46% yield). LCMS RT=1.16 min,ES+ve m/z 375.2/377.1 [M+H]⁺.

15-Iodo-1-phenyl-2,5,8,11-tetraoxapentadecane

A mixture of 15-bromo-1-phenyl-2,5,8,11-tetraoxapentadecane (711 mg,1.894 mmol) and sodium iodide (568 mg, 3.79 mmol) in acetone (10 mL) washeated under reflux conditions for 4 hours. The reaction was cooled toroom temperature. The mixture was filtered through celite and the solidwashed with acetone. The solvent was removed under reduced pressure. Theresidue was dissolved in ethyl acetate (30 mL) and washed with water (30mL) and brine (2×30 mL). The organic extract was dried using ahydrophobic frit and concentrated under reduced pressure to afford thetitle compound (759 mg, 1.797 mmol, 95% yield). LCMS RT=1.23 min, ES+vem/z 440.0 [M+NH₄]⁺.

(7S,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7-(1-phenyl-2,5,8,11-tetraoxapentadecan-15-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one

A solution of KOtBu in THF (1M, 1.282 mL, 1.282 mmol) was added to acooled solution (0° C.) of(8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(240 mg, 0.641 mmol) in anhydrous THF (2 mL). The reaction mixture wasstirred at 0° C. for 45 minutes and then cooled to −78° C. A solution of15-iodo-1-phenyl-2,5,8,11-tetraoxapentadecane (789 mg, 1.868 mmol) inTHF (1 mL) was added dropwise. The solution was stirred at −78° C. for 2minutes, allowed to warm to 0° C. and stirred for 1.5 hours at thattemperature. The reaction was partitioned between water (30 mL) andethyl acetate (30 mL). The organic extract was dried using a hydrophobicfrit and concentrated under reduced pressure. The product was purifiedby chromatography on silica using a gradient elution from 0% to 100%ethyl acetate in cyclohexane to afford the title compound (234 mg, 0.350mmol, 55% yield). LCMS RT=1.48 min, ES+ve m/z 669.3 [M+H]⁺, 686.4[M+NH₄]⁺.

(7S,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7-(1-phenyl-2,5,8,11-tetraoxapentadecan-15-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one

A solution of aqueous HCl (6M, 2.3 mL, 13.80 mmol) was added to asolution of(7S,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7-(1-phenyl-2,5,8,11-tetraoxapentadecan-15-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(234 mg, 0.350 mmol) in THF (2.3 mL). The reaction mixture was stirredat room temperature for 16 hours. Water (30 mL) was added and theproduct was extracted with ethyl acetate (50 mL). The organic extractwas washed with brine (2×30 mL), dried using a hydrophobic frit andconcentrated under reduced pressure to afford the title compound (200mg, 0.344 mmol, 98% yield). LCMS RT=1.07 min, ES+ve m/z 581.3 [M+H]⁺,598.3 [M+NH₄]⁺.

(7R,8R,9S,13S,14S,17S)-13-Methyl-7-(1-phenyl-2,5,8,11-tetraoxapentadecan-15-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol

Triethylsilane (commercially available from for example Aldrich) (0.550mL, 3.44 mmol) was added to a solution of(7S,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7-(1-phenyl-2,5,8,11-tetraoxapentadecan-15-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(200 mg, 0.344 mmol) in TFA (2 mL, 26.0 mmol). The reaction was stirredat room temperature under an atmosphere of nitrogen for 16 hours. Themixture was partitioned between ethyl acetate (30 mL) and brine (30 mL).The organic extract was washed with brine (2×30 mL), dried using ahydrophobic frit and concentrated under reduced pressure. The residuewas dissolved in MeOH (5 mL) and treated with aqueous NaOH (2M, 5 mL,10.00 mmol). The reaction mixture was stirred at room temperature for 3hours. The solvent was removed under reduced pressure. The residue waspartitioned between ethyl acetate (30 mL) and a 10% citric acid solution(30 mL). The organic extract was washed with brine (30 mL), dried usinga hydrophobic frit and concentrated under reduced pressure. The productwas purified by reverse phase chromatography using a gradient elutionfrom 5% to 95% acetonitrile (+0.1% formic acid) in water (+0.1% formicacid) to afford the title compound (150 mg, 0.265 mmol, 77% yield). LCMSRT=1.18 min, ES+ve m/z 567.3 [M+H]⁺, 584.3 [M+NH₄]⁺.

15-((7R,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-1-phenyl-2,5,8,11-tetraoxapentadecane

A vial was charged with(7R,8R,9S,13S,14S,17S)-13-methyl-7-(1-phenyl-2,5,8,11-tetraoxapentadecan-15-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol(150 mg, 0.265 mmol) and DIPEA (0.555 mL, 3.18 mmol) in THF (10 mL). Thevial was sealed, the solution was cooled to 0° C. andchloro(methoxy)methane (commercially available from for example Aldrich)(0.2 mL, 2.63 mmol) was added. The reaction mixture was warmed to roomtemperature, stirred for 1 hour and heated at 70° C. for 40 hours. Thereaction was cooled to room temperature. The reaction was partitionedbetween ethyl acetate (100 mL) and water (100 mL). The organic extractwas washed with brine (2×50 mL), dried using a hydrophobic frit andconcentrated under reduced pressure. The product was purified bychromatography on silica using a gradient elution from 0% to 100% methyltert-butyl ether in cyclohexane to afford the title compound (122 mg,0.186 mmol, 70% yield). LCMS RT=1.60 min, ES+ve m/z 672.5 [M+NH₄]⁺.

2-(2-(2-(4-((7R,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)butoxy)ethoxy)ethoxy)ethanol

A mixture of15-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-1-phenyl-2,5,8,11-tetraoxapentadecane(115 mg, 0.176 mmol) and 10% w/w palladium on carbon (100 mg, 0.094mmol) in ethanol (4 mL) was stirred at room temperature under anatmosphere of hydrogen for 1.5 hours. The palladium on carbon wasfiltered through celite and the filtrate evaporated under reducedpressure. The residue was partitioned between ethyl acetate (15 mL) andbrine (15 mL). The organic extract was dried using a hydrophobic fritand concentrated under reduced pressure to afford the title compound (81mg, 0.143 mmol, 82% yield). LCMS RT=1.36 min, ES+ve m/z 582.4 [M+NH₄]⁺.

Tert-butyl16-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12-tetraoxahexadecan-1-oate

Sodium hydride, 60% w/w in mineral oil (10 mg, 0.250 mmol) was added toa cooled solution (0° C.) of2-(2-(2-(4-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)butoxy)ethoxy)ethoxy)ethanol(81 mg, 0.143 mmol) in DMF (2 mL). The reaction was stirred at thattemperature for 10 minutes and tert-butyl 2-bromoacetate (commerciallyavailable from for example Aldrich) (32 μL, 0.217 mmol) was added. Thereaction was stirred at 0° C. for 1 hour and at room temperature forfurther 2 hours. The reaction mixture was partitioned between ethylacetate (30 mL) and water (30 mL). The organic layer was washed withbrine (30 mL), dried (hydrophobic frit) and concentrated under reducedpressure. The product was purified by chromatography on silica using agradient elution from 0% to 100% methyl tert-butyl ether in cyclohexaneto afford the title compound (60 mg, 0.088 mmol, 62% yield). LCMSRT=1.57 min, ES+ve m/z 696.5 [M+NH₄]⁺.

16-((7R,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12-tetraoxahexadecan-1-oicacid

Tert-butyl16-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12-tetraoxahexadecan-1-oate(133 mg, 0.196 mmol) was dissolved in THF (1.5 mL) and treated withaqueous HCl (6M, 1.5 mL, 9.00 mmol). The reaction mixture was stirred atroom temperature for 8 hours. The reaction mixture was subjecteddirectly to purification by mass-directed automated preparative HPLC(formic acid modifier) to afford the title compound (60 mg, 0.112 mmol,57% yield). LCMS RT=0.89 min, ES+ve m/z 535.3 [M+H]⁺, 552.3 [M+NH₄]⁺.

(2S,4R)-1-((S)-19-((7R,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-2-isopropyl-4-oxo-6,9,12,15-tetraoxa-3-azanonadecan-1-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

HATU (16 mg, 0.042 mmol) was added to a mixture of(2S,4R)-1-((S)-2-amino-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (25 mg, 0.055 mmol),16-((7R,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12-tetraoxahexadecan-1-oicacid (15 mg, 0.028 mmol) and DIPEA (0.05 mL, 0.286 mmol) in DMF (1 mL).The reaction was stirred at room temperature for 30 minutes. Thereaction mixture was subjected directly to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (20 mg, 0.021 mmol, 76% yield). LCMS RT=0.99 min, ES+ve m/z933.3 [M+H]⁺.

(2S,4R)-1-((S)-2-(Tert-butyl)-19-((7R,8R,9S,13S,14S,17S)-3,17-dihydroxy-1-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-4-oxo-6,9,12,15-tetraoxa-3-azanonadecan-1-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

HATU (22 mg, 0.058 mmol) was added to a mixture of(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (25 mg, 0.054 mmol),16-((7R,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12-tetraoxahexadecan-1-oicacid (23 mg, 0.043 mmol) and DIPEA (0.040 mL, 0.229 mmol) in DMF (1 mL).The reaction was stirred at room temperature for 10 minutes. Thereaction mixture was subjected directly to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (26 mg, 0.027 mmol, 64% yield). LCMS RT=1.02 min, ES+ve m/z947.8 [M+H]⁺.

((2-(2-(4-Bromobutoxy)ethoxy)ethoxy)methyl)benzene

To a suspension of sodium hydride, 60% w/w in mineral oil (0.92 g, 22.9mmol) in DMF (5 mL) was added a solution of2-(2-(benzyloxy)ethoxy)ethanol (commercially available from for exampleAldrich) (2.74 mL, 15.29 mmol) in DMF (5 mL) at 0° C. After stirring for25 min, 1,4-dibromobutane (commercially available from for exampleAldrich) (14.9 g, 68.8 mmol) dissolved in DMF (5 mL) was added dropwiseto the mixture. The reaction was warmed to ambient temperature andstirred under an atmosphere of nitrogen for 2.5 hours. A further aliquotof sodium hydride, 60% w/w in mineral oil (0.92 g, 22.9 mmol) was addedand the reaction was stirred at 0° C. for 30 minutes and at ambienttemperature for 30 minutes. A final aliquot of sodium hydride, 60% w/win mineral oil (0.92 g, 22.9 mmol) was added and the reaction stirred atambient temperature for 2 hours then left standing overnight. Thereaction mixture was filtered through celite and the solid washed withDCM. The filtrate was partitioned between DCM (30 mL) and water (30 mL).The organic extract was washed with brine (2×30 mL), dried using ahydrophobic frit, and concentrated under reduced pressure. The productwas purified by chromatography on silica using a using a gradientelution from 0% to 80% methyl tert-butyl ether in cyclohexane to affordthe title compound (3 g, 9.06 mmol, 59% yield). LCMS RT=1.19 min, ES+vem/z 331.2/333.2 [M+H]⁺.

((2-(2-(4-Iodobutoxy)ethoxy)ethoxy)methyl)benzene

A mixture of ((2-(2-(4-bromobutoxy)ethoxy)ethoxy)methyl)benzene (3 g,9.06 mmol) and sodium iodide (2.72 g, 18.11 mmol) in acetone (10 mL) washeated under reflux conditions for 3 hours. The reaction was cooled toroom temperature. The mixture was filtered through celite and the solidwashed with acetone. The solvent was removed under reduced pressure andthe residue was dissolved in ethyl acetate (30 mL) and washed with water(30 mL) and brine (2×30 mL). The organic extract was dried using ahydrophobic frit and concentrated under reduced pressure. The productwas purified by chromatography on silica using a using a gradientelution from 0% to 50% methyl tert-butyl ether in cyclohexane to affordthe title compound (3.1 g, 8.2 mmol, 90% yield). LCMS RT=1.25 min, ES+vem/z 379.2 [M+H]⁺.

(7S,8R,9S,13S,14S,17S)-7-(4-(2-(2-(Benzyloxy)ethoxy)ethoxy)butyl)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one

A solution of KOtBu, in THF (1M, 3.2 mL, 3.2 mmol) was added to a cooledsolution (0° C.) of(8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(600 mg, 1.6 mmol) in anhydrous THF (6 mL). The reaction mixture wasstirred at 0° C. for 45 minutes and then cooled to −78° C. A solution of((2-(2-(4-Iodobutoxy)ethoxy)ethoxy)methyl)benzene (910 mg, 2.4 mmol) inTHF (3 mL) was added dropwise. The solution was stirred at −78° C. for 2minutes then allowed to warm to 0° C. and stirred for 1.5 hours at thattemperature. The reaction was partitioned between water (30 mL) andethyl acetate (30 mL). The organic extract was separated, dried using ahydrophobic frit and concentrated under reduced pressure. The productwas purified by chromatography on silica using a gradient elution from0% to 50% ethyl acetate in cyclohexane to afford the title compound (450mg, 0.72 mmol, 45% yield). LCMS RT=1.49 min, ES+ve m/z 625.5 [M+H]⁺.

(7S,8R,9S,13S,14S,17S)-7-(4-(2-(2-(Benzyloxy)ethoxy)ethoxy)butyl)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one

A solution of aqueous HCl (6M, 4.6 mL, 27.6 mmol) was added to asolution of(7S,8R,9S,13S,14S,17S)-7-(4-(2-(2-(benzyloxy)ethoxy)ethoxy)butyl)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(470 mg, 0.752 mmol) in THF (4.6 mL). The reaction mixture was stirredat room temperature for 18 hours. Water (30 mL) was added and theproduct was extracted with ethyl acetate (50 mL). The organic extractwas washed with brine (2×30 mL), dried using a hydrophobic frit andconcentrated under reduced pressure to afford the title compound (390mg, 0.727 mmol, 97% yield). LCMS RT=1.08 min, ES+ve m/z 537.2 [M+H]⁺,554.2 [M+NH₄]⁺.

(7R,8R,9S,13S,14S,17S)-7-(4-(2-(2-(Benzyloxy)ethoxy)ethoxy)butyl)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol

Triethylsilane (commercially available from for example Aldrich) 1.161mL, 7.27 mmol) was added to a solution of(7S,8R,9S,13S,14S,17S)-7-(4-(2-(2-(benzyloxy)ethoxy)ethoxy)butyl)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(390 mg, 0.727 mmol) in TFA (4.2 mL, 54.5 mmol). The reaction wasstirred at room temperature under an atmosphere of nitrogen for 18hours. The mixture was partitioned between ethyl acetate (50 mL) andbrine (50 mL). The organic extract was washed with brine (2×30 mL),saturated sodium bicarbonate (30 mL), dried using a hydrophobic frit andconcentrated under reduced pressure. The residue was dissolved in MeOH(10 mL) and treated with aqueous NaOH (2M, 5 mL, 10.0 mmol). Thereaction mixture was stirred at room temperature for 1 hour. The solventwas removed under reduced pressure. The residue was partitioned betweenethyl acetate (30 mL) and aqueous HCl solution (1M, 20 mL). The organicextract was washed with brine (20 mL), dried using a hydrophobic fritand concentrated under reduced pressure. The product was purified byreverse phase chromatography using a gradient elution from 5% to 95%acetonitrile (+0.1% formic acid) in water (+0.1% formic acid) to affordthe title compound (270 mg, 0.517 mmol, 71% yield). LCMS RT=1.18 min,ES+ve m/z 523.5 [M+H]⁺, 540.5 [M+NH₄]⁺.

(7R,8R,9S,13S,14S,17S)-7-(4-(2-(2-(Benzyloxy)ethoxy)ethoxy)butyl)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene

Chloro(methoxy)methane (commercially available from for example Aldrich)(0.390 mL, 5.14 mmol) was added to a cooled (0° C.) solution of(7R,8R,9S,13S,14S,17S)-7-(4-(2-(2-(benzyloxy)ethoxy)ethoxy)butyl)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol(270 mg, 0.517 mmol) and DIPEA (1.083 mL, 6.20 mmol) in THF (16 mL). Thereaction mixture was warmed to room temperature, stirred for 1 hour andthen heated at 70° C. for 40 hours. The reaction mixture was cooled to0° C., additional DIPEA (0.271 mL, 1.550 mmol) andchloro(methoxy)methane (0.098 mL, 1.291 mmol) was added. The reactionwas heated to 70° C. and stirred for a further 24 hours. The reactionwas cooled to room temperature, and was partitioned between ethylacetate (100 mL) and water (100 mL). The organic extract was washed withbrine (2×50 mL), dried using a hydrophobic frit and concentrated underreduced pressure. The product was purified by chromatography on silicausing a gradient elution from 0% to 100% methyl tert-butyl ether incyclohexane to afford the title compound (220 mg, 0.36 mmol, 70% yield).LCMS RT=1.62 min, ES+ve m/z 628.6 [M+NH₄]⁺.

2-(2-(4-((7R,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)butoxy)ethoxy)ethanol

A mixture of(7R,8R,9S,13S,14S,17S)-7-(4-(2-(2-(benzyloxy)ethoxy)ethoxy)butyl)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene(220 mg, 0.36 mmol) and 10% w/w palladium on carbon (100 mg, 0.094 mmol)in ethanol (4 mL) was stirred at room temperature under an atmosphere ofhydrogen for 1 hour. The palladium on carbon was filtered throughcelite, washed with ethanol (50 ml) and the filtrate was evaporatedunder reduced pressure to afford the title compound (186 mg, 0.357 mmol,99% yield) LCMS RT=1.36 min, ES+ve m/z 521.5 [M+H]⁺, 538.5 [M+NH₄]⁺.

Tert-butyl2-(2-(2-(4-((7R,8R,9S,13S,14S,17S)-3,17-iso(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)butoxy)ethoxy)ethoxy)acetate

Sodium hydride, 60% w/w mineral oil (25.0 mg, 0.625 mmol) was added to acooled solution (0° C.) of2-(2-(4-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)butoxy)ethoxy)ethanol(186 mg, 0.357 mmol) in DMF (4.5 mL). The reaction was stirred at 0° C.for 10 minutes and tert-butyl 2-bromoacetate (commercially availablefrom for example Aldrich) (79 μL, 0.536 mmol) was added. The reactionwas stirred at 0° C. for 1 hour and at room temperature for a further 6hours. The reaction was cooled to 0° C. and additional sodium hydride,60% w/w in mineral oil (15.72 mg, 0.393 mmol), followed by tert-butyl2-bromoacetate (0.053 mL, 0.357 mmol) was added. The reaction wasstirred at room temperature for a further 18 hours. The reaction mixturewas partitioned between ethyl acetate (30 mL) and water (30 mL). Theorganic layer separated, washed with brine (30 mL), dried using ahydrophobic frit and concentrated under reduced pressure. The productwas purified by chromatography on silica using a gradient elution from0% to 100% methyl tert-butyl ether in cyclohexane to afford the titlecompound (90 mg, 0.142 mmol, 40% yield). LCMS RT=1.56 min, ES+ve m/z652.6 [M+NH₄]+, 657.5 [M+Na]⁺.

2-(2-(2-(4-((7R,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)butoxy)ethoxy)ethoxy)aceticacid

Tert-butyl2-(2-(2-(4-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)butoxy)ethoxy)ethoxy)acetate(80 mg, 0.126 mmol) was dissolved in THF (1 mL) and treated with aqueousHCl (6M, 1 mL, 6.0 mmol). The reaction mixture was stirred at roomtemperature for 4 hours. The reaction mixture was subjected directly topurification by mass-directed automated preparative HPLC (formic acidmodifier) to afford the title compound (23 mg, 0.047 mmol, 37% yield).LCMS RT=0.89 min, ES+ve m/z 491.4 [M+H]⁺.

(2S,4R)-1-((S)-16-((7R,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-2-isopropyl-4-oxo-6,9,12-trioxa-3-azahexadecan-1-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

HATU (12 mg, 0.03 mmol) was added to a mixture of(2S,4R)-1-((S)-2-amino-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (23 mg, 0.05 mmol),2-(2-(2-(4-((7R,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)butoxy)ethoxy)ethoxy)aceticacid (10 mg, 0.02 mmol) and DIPEA (0.04 mL, 0.20 mmol) in DMF (0.8 mL).The reaction was stirred at room temperature for 30 min. The reactionmixture was subjected directly to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (15 mg, 0.017 mmol, 84% yield). LCMS RT=0.98 min, ES+ve m/z889.7 [M+H]⁺.

18-Bromo-1-phenyl-2,5,8,11,14-pentaoxaoctadecane

To a suspension of sodium hydride, 60% w/w in mineral oil (0.85 g, 21.3mmol) in DMF (8 mL) was added a solution of1-phenyl-2,5,8,11-tetraoxatridecan-13-ol (commercially available fromfor example TCI Europe Fine Chemicals) (4.0 g, 14.2 mmol) in DMF (8 mL)at 0° C. After stirring for 25 minutes, 1,4-dibromobutane (commerciallyavailable from for example Aldrich) (7.62 mL, 63.8 mmol) dissolved inDMF (8 mL) was added dropwise to the mixture. The reaction was warmed toroom temperature and stirred under an atmosphere of nitrogen for 30minutes. A further aliquot of sodium hydride, 60% w/w in mineral oil(0.85 g, 21.3 mmol) was added and the reaction was stirred at roomtemperature overnight. Another aliquot of sodium hydride, 60% w/w inmineral oil (0.85 g, 21.3 mmol) was added and the reaction stirred atroom temperature for 2 hours. A final aliquot of sodium hydride, 60% w/win mineral oil (0.43 g, 10.6 mmol) was added and the reaction stirred atroom temperature for 1 hour. The reaction mixture was filtered throughcelite and the solid washed with DCM. The filtrate was partitionedbetween DCM (50 mL) and water (50 mL). The organic extract was washedwith brine (2×50 mL), dried using a hydrophobic frit and concentratedunder reduced pressure. The product was purified by chromatography onsilica using a gradient elution from 0% to 85% methyl tert-butyl etherin cyclohexane to afford the title compound (3.93 g, 9.37 mmol, 63%yield). LCMS RT=1.16 min, ES+ve m/z 419.3/421.2 [M+H]⁺.

18-Iodo-1-phenyl-2,5,8,11,14-pentaoxaoctadecane

A mixture of 18-bromo-1-phenyl-2,5,8,11,14-pentaoxaoctadecane (2.08 g,4.91 mmol) and sodium iodide (1.47 g, 9.82 mmol) in acetone (10 mL) washeated under reflux conditions for 3 hours. The reaction was cooled toroom temperature, filtered through celite and the solid was washed withacetone. The solvent was removed under reduced pressure. The residue wasdissolved in ethyl acetate (30 mL), and washed with water (30 mL) andbrine (2×30 mL). The organic extract was dried using a hydrophobic fritand concentrated under reduced pressure to afford the title compound(759 mg, 1.80 mmol, 95% yield). LCMS RT=1.21 min, ES+ve m/z 467.0[M+H]⁺, 484.0 [M+NH₄]⁺.

(7S,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7-(1-phenyl-2,5,8,11,14-pentaoxaoctadecan-18-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one

A solution of KOtBu in THF (1M, 5.34 mL, 5.34 mmol) was added to acooled solution (0° C.) of(8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(1 g, 2.67 mmol) in anhydrous THF (10 mL). The reaction mixture wasstirred at 0° C. for 45 minutes and then cooled to −78° C.18-Iodo-1-phenyl-2,5,8,11,14-pentaoxaoctadecane (1.87 g, 4.01 mmol) inTHF (5 mL) was added dropwise. The solution was stirred at −78° C. for 2minutes, allowed to warm to 0° C. and stirred for 1.5 hours at thattemperature. The reaction was partitioned between water (50 mL) andethyl acetate (2×50 mL). The organic extracts were dried using ahydrophobic frit and concentrated under reduced pressure. The productwas purified by chromatography on silica using a gradient elution from0% to 100% ethyl acetate in cyclohexane to afford the title compound(883 mg, 1.24 mmol, 46% yield). LCMS RT=1.47 min, ES+ve m/z 713.5[M+H]⁺.

(7S,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7-(1-phenyl-2,5,8,11,14-pentaoxaoctadecan-18-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one

A solution of aqueous HCl (6M, 9.2 mL, 55.2 mmol) was added to asolution of(7S,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7-(1-phenyl-2,5,8,11,14-pentaoxaoctadecan-18-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(883 mg, 1.24 mmol) in THF (9.2 mL). The reaction mixture was stirred atroom temperature for 18 hours. Water (30 mL) was added and the productwas extracted with ethyl acetate (50 mL). The organic extract was washedwith brine (2×30 mL), dried using a hydrophobic frit and concentratedunder reduced pressure to afford the title compound (772 mg, 1.23 mmol,99% yield). LCMS RT=1.06 min, ES+ve m/z 625.3 [M+H]⁺, 642.3 [M+NH₄]⁺.

(7R,8R,9S,13S,4S,17S)-3-Methyl-7-(1-phenyl-2,5,8,11,14-pentaoxaoctadecan-18-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol

Triethylsilane (commercially available from for example Aldrich) (2.0mL, 12.9 mmol) was added to a solution of(7S,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7-(1-phenyl-2,5,8,11,14-pentaoxaoctadecan-18-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(830 mg, 1.29 mmol) in TFA (8.5 mL, 110 mmol). The reaction was stirredat room temperature under an atmosphere of nitrogen for 16 hours. Themixture was partitioned between ethyl acetate (50 mL) and brine (50 mL).The organic extract was washed with brine (2×50 mL), saturated sodiumbicarbonate (50 mL), dried using a hydrophobic frit and concentratedunder reduced pressure. The residue was dissolved in MeOH (10 mL) andtreated with aqueous NaOH (2M, 10 mL, 20.0 mmol). The reaction mixturewas stirred at room temperature for 1 hour. The solvent was removedunder reduced pressure. The residue was partitioned between ethylacetate (40 mL) and 1M HCl solution (20 mL). The organic extract waswashed with brine (20 mL), dried using a hydrophobic frit andconcentrated under reduced pressure. The product was purified by reversephase chromatography using a gradient elution from 5% to 90%acetonitrile (+0.1% formic acid) in water (+0.1% formic acid) to affordthe title compound (375 mg, 0.614 mmol, 47% yield). LCMS RT=1.17 min,ES+ve m/z 611.5 [M+H]⁺, 628.6 [M+NH₄]⁺.

18-((7R,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-1-phenyl-2,5,8,11,14-pentaoxaoctadecane

Chloro(methoxy)methane (commercially available from for example Aldrich)(0.5 mL, 6.58 mmol) was added to a cooled (0° C.) solution of(7R,8R,9S,13S,14S,17S)-13-methyl-7-(1-phenyl-2,5,8,11,14-pentaoxaoctadecan-18-yl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol(375 mg, 0.614 mmol) and DIPEA (1.5 mL, 8.59 mmol) in THF (20 mL). Thereaction mixture was warmed to room temperature, stirred for 1 hour andheated at 70° C. for 72 hours. The reaction was cooled to roomtemperature. The reaction was partitioned between ethyl acetate (100 mL)and water (100 mL). The organic extract was washed with brine (2×50 mL),dried using a hydrophobic frit and concentrated under reduced pressure.The product was purified by chromatography on silica using a gradientelution from 0% to 100% methyl tert-butyl ether in cyclohexane to affordthe title compound (357 mg, 0.51 mmol, 72% yield). LCMS RT=1.60 min,ES+ve m/z 716.7 [M+NH₄]+, 721.7 [M+Na]⁺.

16-((7R,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12-tetraoxahexadecan-1-ol

A mixture of18-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-1-phenyl-2,5,8,11,14-pentaoxaoctadecane(357 mg, 0.444 mmol) and 10% w/w palladium on carbon (157 mg, 0.148mmol) in ethanol (5 mL) was stirred at room temperature under anatmosphere of hydrogen for 1.5 hours. The palladium on carbon wasfiltered through celite and the filtrate evaporated under reducedpressure to afford the title compound (300 mg, 0.41 mmol, 93% yield)LCMS RT=1.37 min, ES+ve m/z 609.6 [M+H]⁺, 631.6 [M+Na]⁺.

Tert-butyl19-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12,15-pentaoxanonadecan-1-oate

Sodium hydride, 60% w/w in mineral oil (30 mg, 0.75 mmol) was added to acooled solution (0° C.) of16-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12-tetraoxahexadecan-1-ol(300 mg, 0.43 mmol) in DMF (5 mL). The reaction was stirred at thattemperature for 10 minutes and tert-butyl 2-bromoacetate (0.095 mL,0.643 mmol) was added. The reaction was stirred at 0° C. for 1 hour andthen at room temperature for a further 18 hours. The reaction mixturewas partitioned between ethyl acetate (30 mL) and water (30 mL). Thewater layer was extracted with additional ethyl acetate (2×30 mL), andthe combined organic layers were washed with brine (2×30 mL), dried(hydrophobic frit) and concentrated under reduced pressure. The productwas purified by chromatography on silica using a gradient elution from0% to 100% methyl tert-butyl ether in cyclohexane to afford the titlecompound (177 mg, 0.245 mmol, 57% yield). LCMS RT=1.58 min, ES+ve m/z740.6 [M+NH₄]+, 745.6 [M+Na]⁺.

19-((7R,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12,15-pentaoxanonadecan-1-oicacid

Tert-butyl19-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12,15-pentaoxanonadecan-1-oate(177 mg, 0.189 mmol) was dissolved in THF (2 mL) and treated withaqueous HCl (6M, 2 mL, 12.0 mmol). The reaction mixture was stirred atroom temperature for 7 hours. The reaction mixture was subjecteddirectly to purification by mass-directed automated preparative HPLC(formic acid modifier) to afford the title compound (64 mg, 0.111 mmol,59% yield). LCMS RT=0.92 min, ES+ve m/z 579.4 [M+H]⁺, 596.5 [M+NH₄]⁺.

(2S,4R)-1-((S)-22-((7R,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-2-isopropyl-4-oxo-6,9,12,15,18-pentaoxa-3-azadocosan-1-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

HATU (16 mg, 0.04 mmol) was added to a mixture of(2S,4R)-1-((S)-2-amino-3-methylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (25 mg, 0.06 mmol),19-((7R,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12,15-pentaoxanonadecan-1-oicacid (16 mg, 0.03 mmol) and DIPEA (0.048 mL, 0.28 mmol) in DMF (0.8 mL).The reaction was stirred at room temperature for 30 minutes. Thereaction mixture was subjected directly to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (17.7 mg, 0.018 mmol, 65% yield). LCMS RT=0.99 min, ES+ve m/z977.4 [M+H]⁺.

(2S,4R)-1-((S)-2-(Tert-butyl)-22-((7R,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-4-oxo-6,9,12,15,18-pentaoxa-3-azadocosan-1-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

HATU (16 mg, 0.04 mmol) was added to a mixture of(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (25 mg, 0.05 mmol),19-((7R,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-3,6,9,12,15-pentaoxanonadecan-1-oicacid (16 mg, 0.03 mmol) and DIPEA (0.048 mL, 0.28 mmol) in DMF (0.8 mL).The reaction was stirred at room temperature for 30 minutes. Thereaction mixture was subjected directly to purification by mass-directedautomated preparative HPLC (formic acid modifier) to afford the titlecompound (17.5 mg, 0.017 mmol, 63% yield). LCMS RT=1.03 min, ES+ve m/z991.4 [M+H]⁺.

(7S,8R,9S,13S,14S,17S)-7-(5-(Benzyloxy)pentyl)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one

A solution of KOtBu in THF (1M, 4.81 mL, 4.81 mmol) was added to acooled solution (0° C.) of(8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(900 mg, 2.403 mmol) in anhydrous THF (10 mL). The reaction mixture wasstirred at 0° C. for 45 minutes and then cooled to −78° C.(((5-Iodopentyl)oxy)methyl)benzene (can be prepared following theprocedure described in J. Chem. Soc., Perkin Trans. 1 1990, 129-132)(2.193 g, 7.21 mmol) in THF (0.5 mL) was added dropwise. The solutionwas stirred at −78° C. for 2 minutes and allowed to warm to roomtemperature and stirred for 1 hour at that temperature. The reaction waspartitioned between water (70 mL) and ethyl acetate (70 mL). The organicextract was dried using a hydrophobic frit and concentrated underreduced pressure. The intermediate was purified by chromatography onsilica using a gradient elution from 0% to 50% ethyl acetate incyclohexane. The residue was dissolved in THF (16 mL) and aqueous HCl(6M, 16 mL, 96 mmol) was added. The reaction was stirred at roomtemperature for 16 hours. The reaction mixture was partitioned betweenethyl acetate (20 mL) and water (20 mL). The organic extract was dried(hydrophobic frit) and concentrated under reduced pressure. The productwas purified by reverse phase chromatography using a gradient elutionfrom 5% to 85% acetonitrile (+0.1% formic acid) in water (+0.1% formicacid) to afford the title compound (487 mg, 1.053 mmol, 44% yield). LCMSRT=1.16 min, ES+ve m/z 463.4 [M+H]⁺.

(7R,8R,9S,13S,14S,17S)-7-(5-(Benzyloxy)pentyl)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol

Triethylsilane (1.681 mL, 10.53 mmol) was added to a solution of(7S,8R,9S,13S,14S,17S)-7-(5-(benzyloxy)pentyl)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-6-one(487 mg, 1.053 mmol) in TFA (6 mL, 78 mmol). The reaction was stirred atroom temperature under an atmosphere of nitrogen for 16 hours. Themixture was partitioned between ethyl acetate (30 mL) and brine (30 mL).The organic extract was washed with brine (2×30 mL), dried using ahydrophobic frit and concentrated under reduced pressure. The residuewas dissolved in MeOH (4 mL) and treated with aqueous NaOH (2M, 4 mL,8.00 mmol). The reaction mixture was stirred at room temperature for 3hours. The solvent was removed under reduced pressure. The residue waspartitioned between ethyl acetate (30 mL) and water (30 mL). The organicextract was washed with brine (30 mL), dried (hydrophobic frit) andconcentrated under reduced pressure. The product was purified by reversephase chromatography using a gradient elution from 10% to 95%acetonitrile (+0.1% formic acid) in water (+0.1% formic acid) to affordthe title compound (410 mg, 0.914 mmol, 87% yield). LCMS RT=1.30 min,ES+ve m/z 449.1 [M+H]⁺.

(7R,8R,9S,13S,14S,17S)-7-(5-(Benzyloxy)pentyl)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene

Chloro(methoxy)methane (0.7 mL, 9.22 mmol) was added to solution of(7R,8R,9S,13S,14S,17S)-7-(5-(benzyloxy)pentyl)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3,17-diol(410 mg, 0.914 mmol) and DIPEA (2 mL, 11.45 mmol) in THF (8 mL). Thereaction vessel was sealed, placed under an atmosphere of nitrogen andheated at 70° C. for 2 days. The reaction was cooled to roomtemperature. The reaction was partitioned between ethyl acetate (50 mL)and brine (50 mL). The organic extract was washed with brine (2×50 mL),dried using a hydrophobic frit and concentrated under reduced pressure.The product was purified by chromatography on silica using a gradientelution from 0% to 100% methyl tert-butyl ether in cyclohexane to affordthe title compound (474 mg, 0.883 mmol, 97% yield). LCMS RT=1.72 min,ES+ve m/z 554.5 [M+NH₄]⁺.

5-((7R,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)pentan-1-ol

A mixture of(7R,8R,9S,13S,14S,17S)-7-(5-(benzyloxy)pentyl)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene(474 mg, 0.883 mmol) and 10% w/w palladium on carbon (100 mg, 0.094mmol) in ethanol (5 mL) and methyl tert-butyl ether (2 mL) was stirredat room temperature under an atmosphere of hydrogen for 1.5 hours. Thepalladium was filtered through celite and the filtrate concentratedunder reduced pressure to afford the title compound (371 mg, 0.831 mmol,94% yield). LCMS RT=1.39 min, ES+ve m/z 447.5 [M+H]⁺ (weak ionisation).

5-((7R,8R,9S,13S,14S,17S)-3,17-Bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)pentyl4-methylbenzenesulfonate

4-Methylbenzene-1-sulfonyl chloride (400 mg, 2.098 mmol) was added to5-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)pentan-1-ol(371 mg, 0.831 mmol) in pyridine (5 mL). The reaction mixture wasstirred at room temperature for 20 hours. The reaction mixture waspartitioned between ethyl acetate (30 mL) and aqueous HCl (2M, 30 mL).The organic extract was washed with sat Na₂CO₃ (30 mL), brine (30 mL),dried (hydrophobic frit) and concentrated under reduced pressure. Theproduct was purified by chromatography on silica using a gradientelution from 0% to 100% methyl tert-butyl ether in cyclohexane to affordthe title compound (401 mg, 0.667 mmol, 80% yield). LCMS RT=1.60 min,ES+ve m/z 623.4 [M+Na]⁺.

Tert-butyl18-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-13-methyl-4,7,10-trioxa-13-azaoctadecan-1-oate,formic acid salt

A microwave vial was charged with5-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)pentyl4-methylbenzenesulfonate (100 mg, 0.166 mmol), tert-butyl5,8,11-trioxa-2-azatetradecan-14-oate (can be prepared following theprocedure described in WO2012054110A2) (145 mg, 0.499 mmol) and DIPEA(0.291 mL, 1.664 mmol) in THF (2 mL). The vial was sealed and placedunder an atmosphere of nitrogen using a vacuum purge. The reaction washeated at 75° C. for 48 hours. The reaction was cooled to roomtemperature. The reaction mixture was subjected directly to purificationby mass-directed automated preparative HPLC (formic acid modifier) toafford the title compound (102 mg, 0.133 mmol, 80% yield). LCMS RT=1.22min, ES+ve m/z 720.6 [M+H]⁺.

18-((7R,8R,9S,13S,14S,17S)-3,17-Dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-13-methyl-4,7,10-trioxa-13-azaoctadecan-1-oicacid, formic acid salt

Tert-butyl18-((7R,8R,9S,13S,14S,17S)-3,17-bis(methoxymethoxy)-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-13-methyl-4,7,10-trioxa-13-azaoctadecan-1-oate,formic acid salt (100 mg, 0.131 mmol) was dissolved in THF (1 mL) andtreated with aqueous HCl (6M, 1 mL, 6.00 mmol). The reaction was stirredat room temperature for 6 hours. The reaction mixture was subjecteddirectly to purification by mass-directed automated preparative HPLC(formic acid modifier) to afford the title compound (24 mg, 0.039 mmol,30% yield). LCMS RT=0.74 min, ES+ve m/z 576.5 [M+H]⁺.

(2S,4R)-1-((S)-2-(Tert-butyl)-21-((7R,8R_(S),13S,14S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-16-methyl-4-oxo-7,10,13-trioxa-3,16-diazahenicosan-1-oyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

HATU (13 mg, 0.034 mmol) was added to a mixture of(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide,hydrochloride (13 mg, 0.028 mmol),18-((7R,8R,9S,13S,14S,17S)-3,17-dihydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-7-yl)-13-methyl-4,7,10-trioxa-13-azaoctadecan-1-oicacid, formic acid salt (12 mg, 0.019 mmol) and DIPEA (0.03 mL, 0.172mmol) in DMF (0.8 mL). The reaction was stirred at room temperature for10 minutes. The reaction mixture was subjected directly to twopurifications by mass-directed automated preparative HPLC (formic acidmodifier followed by ammonium carbonate modifier) to afford the titlecompound (13 mg, 0.013 mmol, 68% yield). LCMS RT=0.84 min, ES+ve m/z988.8 [M+H]⁺.

What is claimed is:
 1. A compound of chemical structure: ULM-L-PTM, wherein: L is a bond or a chemical linker coupling the ULM to the PTM; PTM is a moiety that binds to a target protein or polypeptide which is to be ubiquiniated by VHL; and ULM is a small molecule Von Hippel Lindau E3 ubiquitin ligase (VHL) binding moiety of formula:

wherein R^(1′) is —OH or a group which can be metabolized to —OH; R^(2′) is selected from the group consisting of optionally substituted —NR₁—X^(R2′)-alkyl group, optionally substituted —NR₁—X^(R2′)-Aryl group; optionally substituted —NR₁—X^(R2′)-HET group, optionally substituted —NR₁—X^(R2′)-Aryl-HET group, and optionally substituted —NR₁—X^(R2′)-HET-Aryl group; R₁ is H or C₁-C₃ alkyl; X^(R2′) is optionally substituted —(CH₂)_(n)—, —(CH₂)_(n)—C(X_(v))═C(X_(v))— (cis or trans), —(CH₂)_(n)—C≡C—, —(CH₂CH₂O)_(n)— or C₃-C₆ cycloalkyl; R^(3′) is optionally substituted C₁-C₆ alkyl, optionally substituted —(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)—R^(S3′) group, optionally substituted —(CH₂)_(n)—N(R_(1′))(C═O)_(m′)—(V)_(n′)—R^(S3′) group, optionally substituted —X^(R3′)-alkyl group, optionally substituted —X^(R3′)-Aryl group, optionally substituted —X^(R3′)-HET group, optionally substituted —X^(R3′)-Aryl-HET group, or optionally substituted —X^(R3′)-HET-Aryl group; R^(S3′) is optionally substituted C₁-C₆ alkyl, optionally substituted Aryl group, or a HET group; R_(1′) is H or C₁-C₃ alkyl; V is O, S, or NR_(1′); X^(R3′) is optionally substituted —(CH₂)_(n)—, —(CH₂)_(n)—CH(X_(v))═CH(X_(v))— (cis or trans), —(CH₂)_(n)—CH≡CH—, —(CH₂CH₂O)_(n)— or C₃-C₆ cycloalkyl; X_(v) is H, halo or C₁-C₃ alkyl which is optionally substituted with one or two hydroxyl groups or up to three halogen groups; Alkyl is optionally substituted C₁-C₆ alkyl; Aryl is optionally substituted phenyl or naphthyl; HET is optionally substituted oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, benzofuran, indole, indolizine, azaindolizine, quinoline or a group of chemical structure:

wherein S^(c) is CHR^(SS), NR^(URE), or O; R^(HET) is H, CN, NO₂, halo, optionally substituted C₁-C₆ alkyl, optionally substituted O(C₁-C₆ alkyl) or optionally substituted acetylenic group —C≡C—R_(a), where R_(a) is H or C₁-C₆ alkyl; R^(SS) is H, CN, NO₂, halo, optionally substituted C₁-C₆ alkyl, optionally substituted O—(C₁-C₆ alkyl) or optionally substituted —C(O)(C₁-C₆ alkyl); R^(URE) is H, C₁-C₆ alkyl, or a —C(O)(C₁-C₆ alkyl), each of which groups is optionally substituted with one or two hydroxyl groups or up to three halogens, or optionally substituted heterocycle; and Y^(C) is N or C—R^(YC), where R^(YC) is H, OH, CN, NO₂, halo, optionally substituted C₁-C₆ alkyl, optionally substituted O(C₁-C₆ alkyl) or optionally substituted —C≡C≡C—R_(a) group, where R_(a) is defined as above; R^(PRO) is H, optionally substituted C₁-C₆ alkyl, optionally substituted aryl, optionally substituted heteroaryl or heterocyclic group selected from the group consisting of oxazole, isoxazole, thiazole, isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, quinoline, benzofuran, indole, indolizine, azaindolizine; R^(PRO1) and R^(PRO2) are each independently H, optionally substituted C₁-C₃ alkyl, or together form a keto group; m′ is 0 or 1; each n is independently 0, 1, 2, 3, 4, 5, or 6; each n′ is independently 0 or 1; or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate, or polymorph thereof.
 2. The compound of claim 1, wherein ULM is a group of chemical structure:

wherein R^(1′) is —OH or a group which can be metabolized in a patient or subject to —OH; R^(2′) is a —NR₁—X^(R2′)-Aryl or —NR₁—X^(R2′)-Aryl-HET, wherein Aryl and HET are independently optionally substituted with at least one selected from the group consisting of alkyl, alkoxy, halogen, acid, ester, cycloalkyl and heterocycloalkyl; X^(R2′) is optionally substituted —(CH₂)n; R^(3′) is optionally substituted alkyl, —(CH)R^(CR3′)—NH—C(O)—R^(3P1) or —(CH)R^(CR3′)—R^(3P2)group; R^(CR3′) is optionally substituted C₁-C₄ alkyl; R^(3P1) is optionally substituted C₁-C₆ alkyl, optionally substituted oxetane group, —(CH₂)_(n)OCH₃ wherein n is 1 or 2, or optionally substituted phenyl group, or a morpholino group linked to the carbonyl at the 2- or 3-position; R^(3P2) is

or optionally substituted aryl; HET is optionally substituted thiazole, oxazole, isoxazole, isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrrolidine, furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine, morpholine, benzofuran, indole, indolizine, azaindolizine, and quinolone, wherein the substitution is independently a C1-C3 alkyl group, or a halo group; R^(HET) is H, halo, CN, optionally substituted C₁-C₆ alkyl, optionally substituted C₁-C₆ alkoxy, or optionally substituted aryl; and the ULM group is covalently bonded to a linker group to which is attached a PTM group, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
 3. The compound of claim 2, wherein the ULM is selected from the group consisting of:

wherein the ULM group is covalently bonded to a linker group to which is attached a PTM group, or a pharmaceutically acceptable salt, stereoisomer, solvate, or polymorph thereof.
 4. The compound of claim 1, wherein the ULM is

wherein X is Cl, F, C₁-C₃ alkyl, or an optionally substituted heterocycle; R¹ and R² are each independently H, C₁-C₃ alkyl, or phenyl; and the ULM group is substituted with a linker group or a linker group to which is optionally attached a PTM group, or a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate, or polymorph thereof.
 5. The compound of claim 1, wherein the ULM group is:

wherein n is 0 or 1; R is a linker or a linker attached to a PTM group linked to the ULM group through an amide, ester, ether, carbamate or heterocyclic group; and X is independently is H, F, Cl, C₁-C₃ alkyl optionally substituted with one or two hydroxyl groups, heterocyle, —O—C(O)NR³R⁴ or —C(O)NR³R⁴, wherein each of R³ and R⁴ is independently H, C₁-C₃ alkyl, or phenyl, or a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate or polymorph thereof.
 6. The compound of claim 1, wherein L is at least one member selected from the group consisting of a bond, —(CH₂)_(i)—O—, —(CH₂—CH₂—O)_(i)—, —(CH₂)_(i)—S—, —(CH₂)_(i)—NR—, —(CH₂)_(i)—X₁Y₁—,

and any combinations thereof, wherein each i is independently 0 to 100; R is H, C₁-C₃ alkyl, an alkanol group, or a heterocyclic group; Y is independently a bond, O, S or N—R; X₁Y₁ forms an amide group, a urethane group, ester or thioester group; D is independently a bond (absent),

j is an integer ranging from 1 to 100; k is an integer ranging from 1 to 100; m′ is an integer ranging from 1 to 100; n is an integer ranging from 1 to 100; and X¹ is O, S or N—R; and CON is a bond (absent), a piperazinyl group, optionally substituted alkylene, heterocycle,

X² is O, S, NR⁴, S(O), S(O)₂, —S(O)₂O, —OS(O)₂, or OS(O)₂O; X³ is O, S, CHR⁴ or NR⁴; and R⁴ is H or a C₁-C₃ alkyl group, which is optionally substituted with one or two hydroxyl groups.
 7. The compound of claim 6, wherein i is 1 to
 10. 8. The compound of claim 6, wherein L is a (poly)ethyleneglycol having from 1 to 100 ethylene glycol units.
 9. The compound of claim 6, wherein R is morpholino, piperidinyl, or piperazinyl.
 10. The compound of claim 6, wherein CON is a

group, an amide group, or a piperazinyl group.
 11. The compound of claim 1, wherein the ULM is selected from the group consisting of:

wherein the ULM is modified to be covalently bonded to the PTM group through a linker group, or a pharmaceutically acceptable salt, enantiomer, diasteromer, solvate, or polymorph thereof.
 12. The compound of claim 1, wherein the ULM is a member selected from the group consisting of:

wherein the ULM is modified to be covalently bonded to the PTM group through a linker group, or a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate, polymorph or prodrug thereof.
 13. A compound selected from the group consisting of

wherein each of R_(1PC), R_(2PC), R_(3PC), R_(4PC), R_(5PC), R_(6PC), R_(7PC), R_(8PC), R_(9PC), R_(10PC), R_(11PC), R_(12PC), R_(13PC) and R_(14PC) is are each independently H or a

group; and wherein L is a linker group and PTM is a protein targeting moiety, or a pharmaceutically acceptable salt, enantiomer, diasteromer, solvate, or polymorph thereof.
 14. The compound of claim 13, which is:

wherein R_(7PC) and R_(10PC) are each independently a

group or H, or a pharmaceutically acceptable salt, enantiomer, diasteromer, solvate, or polymorph thereof.
 15. The compound of claim 13, which is:

wherein R_(7PC), R_(11PC) R_(12PC), R_(13PC) and R_(14PC) are each independently a

group or H, or a pharmaceutically acceptable salt, enantiomer, diasteromer, solvate, or polymorph thereof.
 16. The compound of claim 13, which is:

wherein R_(7PC), R_(8PC), R_(9PC), R_(10PC) are R each independently a

group or H, or a pharmaceutically acceptable salt, enantiomer, diasteromer, solvate, or polymorph thereof.
 17. The compound of claim 1, wherein the linker group is a polyethylene group comprising from 1 to 10 ethylene glycol units.
 18. The compound of claim 1, wherein the PTM group is a moiety which binds to a target protein selected from the group consisting of B7.1 and B7, TINFRlm, TNFR2, NADPH oxidase, Bcl, Bax, C5a receptor, HMG-CoA reductase, PDE V phosphodiesterase, PDE IV phosphodiesterase, PDE I, PDEII, PDEIII, squalene cyclase inhibitor, CXCR1, CXCR2, nitric oxide (NO) synthase, cyclo-oxygenase 1, cyclo-oxygenase 2, 5HT receptor, dopamine receptor, G Protein, Gq, histamine receptor, 5-lipoxygenase, tryptase serine protease, thymidylate synthase, purine nucleoside phosphorylase, GAPDH trypanosomal, glycogen phosphorylase, Carbonic anhydrase, chemokine receptor, JAK, STAT, RXR, H1V 1 protease, HIV 1 integrase, influenza, neuramimidase, hepatitis B reverse transcriptase, sodium channel, multi drug resistance (MDR), protein P-glycoprotein (and MRP), tyrosine kinase, CD23, CD124, tyrosine kinase p56 Ick, CD4, CD5, IL-2 receptor, IL-1 receptor, TNF-alphaR, ICAM1, Cat+ channel, VCAM, VLA-4 integrin, selectin, CD40/CD40L, newokinin receptor, inosine monophosphate dehydrogenase, p38 MAP Kinase, Ras, Raf, MEK, ERK, interleukin-1 converting enzyme, caspase, HCV, NS3 protease, HCV NS3 RNA helicase, glycinamide ribonucleotide formyl transferase, rhinovirus 3C protease, herpes simplex virus-1 (HSV-I), protease, cytomegalovirus (CMV) protease, poly (ADP-ribose) polymerase, cyclin dependent kinase, PI3-kinase, focal adhesion kinase, vascular endothelial growth factor, oxytocin receptor, microsomal transfer protein 5 alpha reductase, angiotensin 11, glycine receptor, noradrenaline reuptake receptor, endothelin receptor, neuropeptide Y receptor, adenosine receptor, adenosine kinase, AMP deaminase, purinergic receptor (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7), farnesyltransferase, geranylgeranyl transferase, TrkA a receptor beta-amyloid, tyrosine kinase Flk-IIKDR, vitronectin receptor, integrin receptor, Her-21 neu, telomerase inhibition, cytosolic phospholipaseA2, EGF receptor tyrosine kinase, IGFR, FKBP ecdysone 20-monooxygenase, GABA gated chloride channel, acetylcholinesterase, voltage-sensitive sodium channel protein, calcium release channel, chloride channel, Acetyl-CoA carboxylase, adenylosuccinate synthetase, protoporphyrinogen oxidase, enolpyruvylshikimate-phosphate synthase.
 19. The compound of claim 1, wherein the PTM group is an Hsp90 inhibitor, a kinase inhibitor, a phosphatase inhibitor, a humanMDM2 inhibitor, a compound which targets human BET Bromodomain-containing proteins, an HDAC inhibitor, a human lysine methyltransferase inhibitor, a compound targeting RAF receptor, a compound targeting FKBP, an angiogenesis inhibitor, an immunosuppressive compound, a compound targeting an aryl hydrocarbon receptor, a compound targeting an androgen receptor, a compound targeting an estrogen receptor, a compound targeting a thyroid hormone receptor, a compound targeting a growth factor receptor, a focal adhesion kinase inhibitor, a compound targeting HIV protease, a compound targeting HIV integrase, a compound targeting HCV protease or a compound targeting acyl protein thioesterase 1 or
 2. 20. The compound of claim 18, wherein the PTM group is selected from the group consisting of: derivatized YKB:

wherein a linker moiety L or a

group is attached via the terminal amide group; derivatized p54:

wherein a linker moiety or a

group is attached via the terminal acetylene group; derivatized (5-[2,4-dihydroxy-5-(1-methylethyl)phenyl]-n-ethyl-4-[4-(morpholin-4-ylmethyl)phenyl]isoxazole-3-carboxamide) having the structure:

wherein a linker moiety L or a

group is attached via the amide group or a hydroxyl group; derivatized PU3:

wherein a linker moiety L or a

group is attached via the butyl group; derivatized Geldanamycin ((4E,6Z,8S,9S,10E,12S,13R,14S,16R)-13-hydroxy-8,14,19-trimethoxy-4,10,12,16-tetramethyl-3,20,22-trioxo-2-azabicyclo[16.3.1] wherein a linker or a

group is attached via the amide group; derivatized tanespimycin(17-alkylamino-17-desmethoxygeldanamycin (17-AAG)) wherein a linker or a

group is attached via the amide group; derivatized 17-(2-dimethyl aminoethyl)amino-17-desmethoxygeldanamycin (17-DMAG), wherein a linker or a

group is attached via the amide group; derivatized erlotinib:

wherein R is a linker moiety L or a

group attached via the ether group; derivatized sunitinib:

wherein R is a linker moiety L or a

group which is attached to the pyrrole moiety; derivatized sorafenib:

wherein R is a linker moiety L or a

group attached to the phenyl moiety; derivatized dasatinib:

wherein R is a linker moiety L or a

group attached to the pyrimidine; derivatized lapatinib:

wherein a linker moiety L or a

group is attached via the terminal methyl of the sulfonyl methyl group; derivatized U09-CX-5279:

wherein a linker moiety L or a

group is attached via the amine (aniline), carboxylic acid or cyclopropyl group; derivatized Y1X:

wherein a linker moiety L or a

group is attached via the propyl group; derivatized Y1W:

wherein a linker moiety L or a

group is attached via the propyl group or the tert-butyl group; derivatized 6TP:

wherein a linker moiety L or a

group is attached via the terminal methyl group bound to amide moiety; derivatized OTP:

wherein a linker moiety L or a

group is attached via the terminal methyl group bound to the amide moiety; derivatized 07U:

wherein a linker moiety L or a

group is attached via the secondary amine or terminal/primary amino group; derivatized YCF:

wherein a linker moiety L or a

group is attached via either of the terminal hydroxyl groups; derivatized XK9:

wherein a linker moiety L or a

group is attached via the terminal hydroxyl group; derivatized NXP:

wherein a linker moiety L or a

group is attached via an amine group; derivatized afatinib, wherein a linker moiety L or a

group is attached via the aliphatic amine group; derivatized fostamatinib, wherein a linker moiety L or a

group is attached via a methoxy group; derivatized gefitinib:

wherein a linker moiety L or a

group is attached via a methoxy or ether group; derivatized lenvatinib, wherein a linker moiety L or a

group is attached via the cyclopropyl group; derivatized vandetanib, wherein a linker moiety L or a

group is attached via the methoxy or hydroxyl group; derivatized vemurafenib, wherein a linker moiety L or a

group is attached via the sulfonyl propyl group; derivatized imatinib:

wherein R is a linker moiety L or a

group attached via the amide group or optionally the L or

group is attached via the aniline amine group; derivatized pazopanib:

wherein R is a linker moiety L or a

group attached to the phenyl moiety or optionally the L or a

group is attached via the aniline amine group; derivatized AT-9283:

wherein R is a linker moiety L or a

group attached to the phenyl moiety; derivatized TAE684:

wherein R is a linker moiety L or a

group which is attached to the phenyl moiety; derivatized nilotinib:

wherein R is a linker moiety L or a

group attached to the phenyl moiety or optionally L or a

group is attached via an aniline amine group; derivatized NVP-BSK805:

wherein R is a linker moiety L or a

group attached to a phenyl moiety or optionally L or a

group is attached via the diazole group; derivatized crizotinib:

wherein R is a linker moiety L or a

group which is attached to the diazole group or optionally L or a

group is attached via a phenyl moiety; derivatized:

wherein R is a linker moiety L or a

group attached to the phenyl moiety; derivatized foretinib:

wherein R is a linker moiety L or a

group which is attached to the phenyl moiety or a hydroxyl or ether group on the quinoline moiety; derivatized PTP1B:

wherein a linker group L or a

group is attached at R; derivatized inhibitor of SHP-2 domain of tyrosine phosphatase:

wherein a linker group L or a

group is attached at R; derivatized inhibitor of BRAF (BRAF^(V600E))/MEK:

wherein a linker L or a

group is attached at R; derivatized inhibitor of tyrosine kinase ABL:

wherein R is a linker group L or a

group; derivatized nutlin-3:

wherein a linker moiety L or a

group is attached at the methoxy group or via a hydroxyl linkage; derivatized nutlin-2:

wherein a linker moiety L or a

group is attached at the methoxy group or via a hydroxyl group;

derivatized nutlin-1: wherein a linker moiety or a

group is attached via the methoxy group or via a hydroxyl group; derivatized trans-4-Iodo-4′-Boranyl-Chalcone, wherein a linker moiety L or a

group is attached via a hydroxy group; a derivatized compound targeting human BET Bromodomain-containing protein Brd2, Brd3, Brd4 selected from the group consisting of:

wherein R is a linker L group or a

group; derivatized SAHA:

wherein R is a linker group L or a

group; derivatized BIX-01294:

wherein each R is independently a linker group L or a

group; derivatized UNC0244:

wherein each R is independently a linker group L or a

group; derivatized azacitidine, wherein a linker group L or a

group is attached via the hydroxy or amino groups; derivatized decitabine, wherein a linker group L or a

group is attached via either of the hydroxy groups or at the amino group; derivatized GA-1; derivatized estradiol; derivatized dihydroxytestosterone; derivatized ovalicin; derivatized fumagillin; derivatized AP21998; a derivatized glucocorticoid selected from the group consisting of hydrocortisone, prednisone, prednisolone, methylprednisoloneand beclometasone dipropionate, wherein a linker group L or a

group is attached; derivatized methotrexate, wherein a linker group L or a

group is attached to either of the terminal hydroxyls; derivatized ciclosporin, wherein a linker group L or a

group is attached at any of the butyl groups; derivatized tacrolimus (FK-506) or rapamycin, wherein a linker group L or a

group is attached at one of the methoxy groups; derivatized actinomycin, wherein a linker group L or a

group is attached at one of the isopropyl groups; a derivatized compound targeting the aryl hydrocarbon receptor (AHR) selected from the group consisting of Apigenin, SR1 and LGC006, wherein a linker group L or a

group is attached; derivatized PLX4032:

wherein R designates a linker group L or a

group; a derivatized compound targeting FKBP according to the chemical structure:

wherein R designates a linker group L or a

group; a derivatized compound targeting androgen receptor (AR) according to the chemical structure:

wherein R designates a linker group L or a

group; a derivatized SARM ligand of Androgen Receptor according to the chemical structure:

wherein R designates a linker group L or a

group; a derivatized Androgen Receptor Ligand DHT according to the chemical structure:

wherein R designates a linker group L or a

group; a derivatized compound targeting Estrogen Receptor (ER) according to the chemical structure:

wherein R designates a linker group L or a

group; a derivatized compound targeting Thyroid Hormone Receptor (TR) according to the chemical structure:

wherein R designates a linker group L or a

group and MOMO indicates a methoxymethoxy group; a derivatized compound targeting HIV Protease according to the chemical structure:

wherein R designates a linker group L or a

group; a derivatized Inhibitor of HIV Integrase according to the chemical structure:

wherein R designates a linker group L or a

group; a derivatized compound targeting HCV Protease according to the chemical structure:

wherein R designates a linker group L or a

group; and a derivatized compound targeting Acyl-protein Thioesterase-1 and -2 (APT1 and APT2) according to the chemical structure:

wherein R designates a linker group L or a

group.
 21. A pharmaceutical composition comprising the compound of claim 1 and at least one pharmaceutically suitable carrier.
 22. A method of regulating the amount or activity of a target protein in a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a compound of claim
 2. 23. The method of claim 22, wherein the target protein is selected from the group consisting of structural proteins, receptors, enzymes, cell surface proteins, aromatases, motor activity proteins, helicases, proteolytic proteins, kinases, oxidoreductases, transferases, hydrolases, lyases, isomerases, ligases, enzyme regulators, signal transduction proteins, lipid binding proteins, carbohydrate binding proteins, receptors, cell motility proteins, membrane fusion proteins, cell communication proteins, developmental proteins, cell differentiation proteins, cell adhesion proteins, pro- and anti-apoptotic proteins, transport proteins, nuclear transport proteins, hormone receptors, ion transporter proteins, carrier proteins, permeases, secretory proteins, electron transport tproteins, chaperone regulator proteins, nucleic acid binding proteins, transcription regulatory proteins, and translation regulator proteins.
 24. The method of claim 22, wherein the target protein is selected from the group consisting of B7.1 and B7, TINFRlm, TNFR2, NADPH oxidase, BclIBax and other partners in the apotosis pathway, C5a receptor, HMG-CoA reductase, PDE V phosphodiesterase type, PDE IV phosphodiesterase type 4, PDE I, PDEII, PDEIII, squalene cyclase inhibitor, CXCR1, CXCR2, nitric oxide (NO) synthase, cyclo-oxygenase 1, cyclo-oxygenase 2, 5HT receptors, dopamine receptors, G Proteins, i.e., Gq, histamine receptors, 5-lipoxygenase, tryptase serine protease, thymidylate synthase, purine nucleoside phosphorylase, GAPDH trypanosomal, glycogen phosphorylase, Carbonic anhydrase, chemokine receptors, JAW STAT, RXR and similar, HIV 1 protease, HIV 1 integrase, influenza, neuramimidase, hepatitis B reverse transcriptase, sodium channel, multi drug resistance (MDR), protein P-glycoprotein (and MRP), tyrosine kinases, CD23, CD124, tyrosine kinase p56 lck, CD4, CD5, IL-2 receptor, IL-1 receptor, TNF-alphaR, ICAM1, Cat+ channels, VCAM, VLA-4 integrin, selectins, CD40/CD40L, newokinins and receptors, inosine monophosphate dehydrogenase, p38 MAP Kinase, RaslRaflMEWERK pathway, interleukin-1 converting enzyme, caspase, HCV, NS3 protease, HCV NS3 RNA helicase, glycinamide ribonucleotide formyl transferase, rhinovirus 3C protease, herpes simplex virus-1 (HSV-I), protease, cytomegalovirus (CMV) protease, poly (ADP-ribose) polymerase, cyclin dependent kinases, vascular endothelial growth factor, oxytocin receptor, microsomal transfer protein inhibitor, bile acid transport inhibitor, 5 alpha reductase inhibitors, angiotensin 11, glycine receptor, noradrenaline reuptake receptor, endothelin receptors, neuropeptide Y and receptor, estrogen receptors, androgen receptors, adenosine receptors, adenosine kinase and AMP deaminase, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7), farnesyltransferases, geranylgeranyl transferase, TrkA a receptor for NGF, beta-amyloid, tyrosine kinase Flk-IIKDR, vitronectin receptor, integrin receptor, Her-21 neu, telomerase inhibition, cytosolic phospholipaseA2 and EGF receptor tyrosine kinase. Additional protein targets include, for example, ecdysone 20-monooxygenase, ion channel of the GABA gated chloride channel, acetylcholinesterase, voltage-sensitive sodium channel protein, calcium release channel, chloride channels, acetyl-CoA carboxylase, adenylosuccinate synthetase, protoporphyrinogen oxidase, and enolpyruvylshikimate-phosphate synthase.
 25. A method of treating a disease state or condition in a patient, wherein the method comprises administering to the subject a therapeutically effective amount of a compound of claim 2, wherein the compound regulates the amount or activity of a target protein in the subject, wherein the target protein's dysregulated amount or activity is responsible for the disease state or condition in the subject.
 26. The method of claim 25, wherein the disease state is cancer.
 27. The method of claim 26, wherein the cancer is squamous-cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas, cancer of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemias; benign and malignant lymphomas, particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; sarcomas, including Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheral neuroepithelioma, synovial sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors, meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowel cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor or teratocarcinomas.
 28. The method of claim 26, wherein the cancer is T-lineage Acute lymphoblastic Leukemia (T-ALL), T-lineage lymphoblastic Lymphoma (T-LL), Peripheral T-cell lymphoma, Adult T-cell Leukemia, Pre-B ALL, Pre-B Lymphomas, Large B-cell Lymphoma, Burkitts Lymphoma, B-cell ALL, Philadelphia chromosome positive ALL, and Philadelphia chromosome positive CML
 29. The method of claim 25, wherein the disease state is prostate cancer, Kennedy's Disease, breast cancer, Lymphoma, diabetes, diabetes melittus type I, diabetes melittus type II, obesity, colorectal cancer, head & neck cancer, immune system disorders, leukemia, stem cell growth, wound healing, atherosclerosis, hepatocellular carcinoma, endometrial cancer, McCune-Albright Syndrome, adenocarcinoma, acute lymphoblastic leukemia (ALL), myeloproliferative diseases, and large B-cell lymphoma.
 30. A method of degrading a target protein in a cell, wherein the method comprises contacting the cell with an effective amount of the compound of claim 1, wherein the compound promotes degradation of the target protein in the cell. 